Aldosterone synthase inhibitors

ABSTRACT

The present invention relates to compounds of formula I: 
                         
and pharmaceutically acceptable salts thereof, wherein R 1 , R 2  and R 3 , are as defined herein. The invention also relates to pharmaceutical compositions comprising these compounds, methods of using these compounds in the treatment of various diseases and disorders, processes for preparing these compounds and intermediates useful in these processes.

FIELD OF THE INVENTION

This invention relates to heteroaryl compounds that are useful asinhibitors of aldosterone synthase (CYP11B2) and are thus useful fortreating a variety of diseases that are mediated or sustained byaldosterone activity, including renal disease, diabetic nephropathy,cardiovascular diseases and fibrotic disorders. This invention alsorelates to pharmaceutical compositions comprising these compounds,methods of using these compounds in the treatment of various diseasesand disorders, processes for preparing these compounds and intermediatesuseful in these processes.

BACKGROUND

Aldosterone is a steroid hormone having mineralcorticoid activity. It isproduced primarily by the adrenal glomerulosa in response to angiotensinII, adrenocorticotropic hormone and increased serum potassium levels. Aprimary physiological role of aldosterone in the kidney is to maintainsodium and potassium balance by regulating cation exchange (Na⁺reabsorption and K⁺ secretion) in the distal nephron. However,aldosterone has also been shown to be a pro-inflammatory and profibrotichormone in blood vessels, heart and kidneys. The effects of aldosteroneon gene expression are mediated via binding to the mineralocorticoidreceptor (MR) and a canonical nuclear hormone receptor pathway. However,the hormone also elicits rapid, non-genomic responses, including acuteregulation of the activity of tubular ion transporters, for exampleNa⁺/H⁺ exchangers (NHEs), H⁺-ATPase, ENaC, and Na⁺/K⁺ ATPase (D. W.Good, 2007, Hypertension, 49, 728-739). It is likely that some of theseeffects are mediated by MR-independent pathways. Conversely, the MR canbind alternative ligands, including deoxycorticosterone, corticosterone,cortisol and progesterone. Thus, inhibition of aldosterone synthesis ispredicted to have a pharmacodynamic profile distinct from what isobserved with MR antagonists.

Aldosterone is synthesized in the zona glomerulosa of the adrenalglands, where a single enzyme, CYP11B2 (aldosterone synthase), catalyzesthe 3-step conversion of 11-deoxycorticosterone (11-DOC) to aldosterone,via corticosterone and 18-hydroxycorticosterone. Adrenal aldosteronesynthase activity is regulated by Angiotensin II and K+ levels andunidentified adipocyte-derived mediators. Low levels of aldosteronesynthase have also been detected in the heart and CNS, though thephysiological relevance is uncertain, perhaps relating to paracrineeffects. Systemic aldosterone is believed to derive essentially entirelyfrom the adrenals.

Beyond its role in regulating sodium and potassium balance, aldosteronehas been shown to have pro-inflammatory and pro-fibrotic actions inmultiple tissues including the kidney, blood vessels and the heart. Theharmful effects of inappropriate aldosterone levels on blood pressureand cardiac, renal, cerebral and vascular function and structure, havebeen widely reported in the literature, including: i) increase in sodiumretention through Na⁺/K⁺ ATPase pump induction in distal tubulesresulting in volume expansion and high blood pressure, ii) endothelialdysfunction, iii) oxidative stress, iv) renal and cardiac hypertrophy,v) fibroblast proliferation, and, vi) excessive synthesis ofextracellular matrix resulting in renal, cardiac and vascular fibrosis.

Benefits of aldosterone blockade/inhibition include reduction of kidneyfibrosis and improvement of glomerular filtration rate and albuminuriain models of chronic kidney disease (CKD) and diabetic nephropathy. Thisis supported by pre-clinical data (for example, Fiebler et al., 2005,Circulation, 111, 3087-3094; Lea et al., 2009, Kidney International, 75,936-945). Other benefits reported in the literature include decreasedblood pressure and end-organ damage (heart, kidney, vessels) in bothrenin-dependent and salt-sensitive hypertension.

Although many of aldosterone's known effects are mediated throughmineralcorticoid receptor (MR) activation, and much of the evidencefavoring targeting this pathway comes from experiments with MRantagonists, non-MR mediated effects are reported and knockout mice forMR and aldosterone synthase exhibit different phenotypes (Makhanova etal. 2006, Berger et al. 1998, Funder 2007). These observations furthersuggest that aldosterone synthase inhibitors may have a differentprofile and offer advantages compared to MR antagonists.

For example, several aldosterone actions are not inhibited by MRantagonists, including the potentially deleterious effects on thevasculature (increased peripheral vascular resistance), the heart(effects on myocardial re-polarization) and the endocrine system(decreased insulin secretion). Furthermore, MR antagonism leads to anincrease in circulating aldosterone, predicted to increase aldosteronesignaling via non-MR pathways and, potentially, partially overcoming theMR blockade itself.

Current therapeutic strategies focus on slowing progression and treatingconditions underlying diabetic nephropathy: control of blood glucose andcontrol of high blood pressure. Angiotensin converting enzyme (ACE)inhibitors and angiotensin receptor blockers (ARB) have shown renalbenefit in diabetic patients. To date, representatives of the ACEinhibitor class and from the ARB class have been approved for thetreatment of diabetic nephropathy. These therapies represent limitedbenefit for the diabetic nephropathy patients.

Although the use of ACE inhibitors and ARBs represents the currentstandard of care for patients with diabetic nephropathy, patientsprogressively lose kidney function while on these medications, as seenin the IDNT (E. J. Lewis et al., 2001, N. Engl. J. Med., 345, 851-860)and RENAAL (B. M. Brenner et al., 2001, N. Engl. J. Med., 345, 861-869)studies, which reported a decrease over time in estimated glomerularfiltration rate, which is an accurate measure of chronic kidney diseaseprogression in patients treated by these conventional methods. At stage5 chronic kidney disease, renal replacement therapy is required, in theform of either dialysis or transplant.

Aldosterone synthase inhibition may also be predicted to offeradvantages as add-on therapy with ACE inhibitors and ARBs. Notably,25-50% of patients receiving these agents experience “aldosteronebreakthrough” in which aldosterone levels initially lowered by thesetreatments eventually return to pretreatment levels. This phenomenonwould not occur with direct aldosterone synthase inhibition and couldenhance efficacy in combination therapy.

There remains a high unmet medical need to treat diabetic nephropathy,to halt or regress disease progression by specifically targeting theunderlying pathophysiological mechanisms associated with chronicinflammation and fibrosis, irrespective of the original cause of thedisease and when co-administered with current therapies. The studiesdescribed above and in the literature provide evidence that inhibitorsof aldosterone synthesis will be useful for the treatment of diabetickidney disease including diabetic nephropathy; non-diabetic kidneydisease including glomerulosclerosis, glomerulonephritis, IGAnephropathy, nephritic syndrome and focal segmental glomerulosclerosis(FSGS); cardiovascular diseases including hypertension, pulmonaryarterial hypertension, Conn's syndrome, systolic heart failure,diastolic heart failure, left ventricular dysfunction, left ventricularstiffness and fibrosis, left ventricular filing abnormalities, arterialstiffness, atherosclerosis and cardiovascular morbidity associated withprimary or secondary hyperaldosteronism; adrenal hyperplasia and primaryand secondary hyperaldosteronism.

BRIEF SUMMARY OF THE INVENTION

The present invention provides novel compounds that inhibit aldosteronesynthase and thus useful for treating a variety of diseases anddisorders that can be alleviated by lowering levels of aldosteroneincluding renal disease, diabetic nephropathy, cardiovascular diseasesand fibrotic disorders. This invention also relates to pharmaceuticalcompositions comprising these compounds, methods of using thesecompounds in the treatment of various diseases and disorders, processesfor preparing these compounds and intermediates useful in theseprocesses.

DETAILED DESCRIPTION OF THE INVENTION

In an embodiment of the invention, there are provided compounds of theformula I

wherein:R¹ is selected from —C(O)NH₂, —C(O)NH(CH₃) and —CN;R² is —(X)—R⁴, wherein—(X)— is a bond, —CH₂—, or —O—; andR⁴ is selected from—H;C₁₋₃alkyl, optionally substituted with one to four groups selected from—F, —OH, and —SO₂C₁₋₃alkyl;halogen;—CN;—SO₂C₁₋₃ alkyl;—C(O)N(C₁₋₃alkyl)₂;—NHC(O)R⁵ or —N(CH₃)C(O)R⁵, provided that —(X)— is —CH₂— and wherein R⁵is selected from C₃₋₆cycloalkyl and C₁₋₃alkyl optionally substitutedwith one to three —F groups;—NHSO₂C₁₋₃alkyl;—CH(cyclopropyl)NHSO₂C₁₋₃ alkyl;—OCH₂C(O)N(C₁₋₃alkyl)₂, provided that —(X)— is —CH₂—;—S(═O)(═NH)CH₃, provided that —(X)— is —CH₂—;heterocyclyl selected from tetrahydropyranyl, tetrahydrofuranyl,pyrrolidinyl, 1,1-dioxo[1,2]-thiazine, morpholinyl, oxazolidinyl,piperidinyl, azetidinyl, wherein said heterocyclyl is optionallysubstituted with one to three groups selected from —C(O)C₁₋₃alkyl,halogen, —OH, oxo and C₁₋₃ alkyl;—C(O)-heterocyclyl, provided that —(X)— is —CH₂, wherein saidheterocyclyl is selected from morpholin-4-yl, pyrrolidin-1-yl andpiperidin-1-yl, optionally substituted with one or two groups selectedfrom —F and —OH;C₃₋₆cycloalkyl optionally substituted with —CN or —OH; andphenyl, optionally substituted with —SO₂NH₂; andR³ is H, or C₁₋₃alkyl optionally substituted with —OH; orR² and R³ together form an annelated five-membered cycloalkyl ringoptionally substituted with —OH;or a salt or a stereoisomer thereof.

In another embodiment, there are provided compounds of the formula I asdescribed according to the embodiment above and wherein

R¹ is —C(O)NH₂ or —CN;

R² is —(X)—R⁴, wherein

—(X)— is a bond, and

R⁴ is selected from

—CH₃;

—CF₃;

—CHF₂;

—CH₂OH;

—CH(OH)CH₃;

—CH(OH)CF₃;

—F;

—CN;

heterocyclyl selected from tetrahydropyranyl and pyrrolidinyl, whereinsaid heterocyclyl is optionally substituted with one to three groupsselected from C₁₋₃alkyl, halogen, —OH and oxo;

C₃₋₆cycloalkyl optionally substituted with —CN or —OH; and phenyl,optionally substituted with —SO₂NH₂; or

—(X)— is O, and

R⁴ is selected from

C₁₋₃alkyl;

—CH₂SO₂C₁₋₃alkyl; and

heterocyclyl selected from tetrahydropyranyl, tetrahydrofuranyl,pyrrolidinyl, piperidinyl, and azetidinyl, wherein said heterocyclyl isoptionally substituted with one to three groups selected from—C(O)C₁₋₃alkyl, halogen, —OH, oxo and C₁₋₃alkyl; or

X is (—CH₂—), and

R⁴ is selected from

—SO₂C₁₋₃alkyl;

—C(O)N(C₁₋₃alkyl)₂;

—NHC(O)R⁵ or —N(CH₃)C(O)R⁵, wherein R⁵ is selected from cyclopropyl andC₁₋₃alkyl optionally substituted with one to three —F groups;

—OCH₂C(O)N(C₁₋₃alkyl)₂;

—NHSO₂C₁₋₃alkyl;

—S(═O)(═NH)CH₃;

heterocyclyl selected from pyrrolidinyl, 1,1-dioxo[1,2]-thiazine,morpholinyl and oxazolidinyl, wherein said heterocyclyl is optionallysubstituted with one to three groups selected from —C(O)C₁₋₃alkyl,halogen, —OH, oxo and C₁₋₃alkyl; and

—C(O)-heterocyclyl, wherein the heterocyclyl is selected frommorpholin-4-yl, pyrrolidin-1-yl and piperidin-1-yl, optionallysubstituted with one or two groups selected from —F and —OH; and

R³ is H or C₁₋₃alkyl optionally substituted with —OH;

or a salt or a stereoisomer thereof.

In another embodiment, there are provided compounds of the formula I asdescribed according to any of the embodiments above and wherein

R² is —(X)—R⁴, wherein

—(X)— is a bond, and

R⁴ is selected from

—CF₃;

—CHF₂;

—CH₂OH;

—CH(OH)CH₃;

—CH(OH)CF₃;

—F;

—CN;

heterocyclyl selected from tetrahydropyranyl and pyrrolidinyl, whereinsaid heterocyclyl is substituted with one to three groups selected fromC₁₋₃alkyl, —F, —OH and oxo;

C₃₋₆cycloalkyl, substituted with —CN or —OH; and

phenyl, optionally substituted with —SO₂NH₂; and

R³ is H, or C₁₋₃alkyl optionally substituted with —OH;

or a salt or a stereoisomer thereof.

In another embodiment, there are provided compounds of the formula I asdescribed according to any of the embodiments above and wherein

R² is —(X)—R⁴, wherein

—(X)— is O, and

R⁴ is selected from

C₁₋₃alkyl;

—CH₂SO₂C₁₋₃alkyl; and

heterocyclyl selected from tetrahydropyranyl, tetrahydrofuranyl,pyrrolidinyl, piperidinyl, and azetidinyl, wherein said heterocyclyl isoptionally substituted with —C(O)C₁₋₃alkyl; and

R³ is H, or C₁₋₃alkyl optionally substituted with —OH;

or a salt or a stereoisomer thereof.

In another embodiment, there are provided compounds of the formula I asdescribed according to any of the embodiments above and wherein

R² is —(X)—R⁴, wherein

X is (—CH₂—), and

R⁴ is selected from

—SO₂C₁₋₃alkyl

—C(O)N(C₁₋₃alkyl)₂;

—NHC(O)R⁵ or —N(CH₃)C(O)R⁵, wherein R⁵ is selected from cyclopropyl andC₁₋₃alkyl optionally substituted with one to three —F groups;

—OCH₂C(O)N(C₁₋₃alkyl)₂;

—NHSO₂C₁₋₃alkyl;

—S(═O)(═NH)CH₃;

heterocyclyl selected from pyrrolidinyl, 1,1-dioxo[1,2]-thiazine,morpholinyl and oxazolidinyl, wherein said heterocyclyl is optionallysubstituted with one to two groups selected from oxo and C₁₋₃alkyl; and

—C(O)-heterocyclyl, wherein the heterocyclyl is selected frommorpholin-4-yl, pyrrolidin-1-yl and piperidin-1-yl, optionallysubstituted with one or two groups selected from —F and —OH; and

R³ is H, or C₁₋₃alkyl optionally substituted with —OH;

or a salt or a stereoisomer thereof.

In another embodiment, there are provided compounds of the formula I asdescribed according to any of the embodiments above and wherein

R¹ is —C(O)NH₂;

or a salt or a stereoisomer thereof.

In another embodiment, there are provided compounds of the formula I asdescribed according to any of the embodiments above and wherein

R¹ is —CN;

or a salt or a stereoisomer thereof.

In another aspect of the invention, there is provided a compound of thegeneral formula I or a stereoisomer or pharmaceutically acceptable saltthereof for use in a therapeutic method as described hereinbefore andhereinafter.

Table 1 shows representative compounds of the invention which can bemade by the methods described in the general synthetic schemes, theexamples, and known methods in the art.

TABLE 1 Cpd Structure Name  1

2-[5-(4-Hydroxy-tetrahydro-pyran-4-yl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine- 5-carboxylic acid amide  2

2-[5-(1-Cyano-cyclopropyl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylic acid amide  3

2-[5-(1,1-Dioxo-1λ⁶,-[1,2]thiazinan-2-ylmethyl)-pyridin-3-yl]-2,3-dihydro- benzo[1,4]dioxine-5-carboxylic acidamide  4

2-Pyridin-3-yl-2,3-dihydro- benzo[1,4]dioxine-5-carboxylic acid amide  5

2-[5-(2-Oxo-pyrrolidin-1-ylmethyl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5- carboxylic acid amide  6

2-[5-(-1-Methyl-5-oxo-pyrrolidin-2-yl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine- 5-carboxylic acid amide  7

2-[5-((R)-1-Acetyl-piperidin-3-yloxy)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine- 5-carboxylic acid amide  8

2-(5-Methanesulfonylmethyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylic acid amide  9

2-(5-Trifluoromethyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylic acid amide 10

2-[5-(Tetrahydro-pyran-4-yl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylic acid amide 11

2-[5-(1-Hydroxy-cyclohexyl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylic acid amide 12

2-[5-(1-Acetyl-piperidin-4-yloxy)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5- carboxylic acid amide 13

2-[5-(2-Morpholin-4-yl-2-oxo-ethyl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5- carboxylic acid amide 14

2-{5-[(Cyclopropanecarbonyl-amino)- methyl]-pyridin-3-yl}-2,3-dihydro-benzo[1,4]dioxine-5-carboxylic acid amide. 15

2-[5-(3-Oxo-morpholin-4-ylmethyl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5- carboxylic acid amide 16

2-[5-(4-Sulfamoyl-phenyl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylic acid amide 17

2-[5-((S)-1-Acetyl-pyrrolidin-3-yloxy)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine- 5-carboxylic acid amide 18

2-[5-((R)-1-Acetyl-pyrrolidin-3-yloxy)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine- 5-carboxylic acid amide 19

2-[5-(1-Acetyl-azetidin-3-yloxy)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5- carboxylic acid amide 20

2-(5-Hydroxymethyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylic acid amide 21

2-(5-Fluoro-4-methyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylic acid amide 22

2-(5-Difluoromethyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxlne-5-carboxylic acid amide 23

2-(4-Methyl-pyridin-3-yl)-2,3-dihydro- benzo[1,4]dioxine-5-carboxylicacid amide 24

2-{5-[(Cyclopropanecarbonyl-methyl-amino)-methyl]-pyridin-3-yl}-2,3-dihydro- benzo[1,4]dioxine-5-carboxylicacid amide 25

2-(5-Dimethylcarbamoylmethoxymethyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine- 5-carboxylic acid amide 26

2-[5-(1-Hydroxy-cyclobutyl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine- 5-carboxylic acid amide 27

2-[5-(1-Hydroxy-ethyl)-4-methyl-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5- carboxylic acid amide 28

2-(5-Trifluoromethyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylic acid methylamide 29

2-[5-[2-(Dimethylamino)-2-oxo-ethyl]-3-pyridyl]-2,3-dihydro-1,4-benzodioxine-5- carboxamide 30

2-{5-[2-(4,4-Difluoro-piperidin-1-yl)-2-oxo-ethyl]-pyridin-3-yl}-2,3-dihydro- benzo[1,4]dioxine-5-carboxylic acidamide 31

2-[5-(2-Oxo-oxazolidin-3-ylmethyl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5- carboxylic acid amide 32

2-[5-(4-Fluoro-tetrahydro-pyran-4-yl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine- 5-carboxylic acid amide 33

2-[5-(1-Acetyl-piperidin-4-yloxy)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5- carboxylic acid methylamide 34

2-[5-(2-Oxo-oxazolidin-3-ylmethyl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5- carbonitrile 35

2-[5-(1-Methyl-5-oxo-pyrrolidin-2-yl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4] dioxine-5-carbonitrile 36

2-[5-(3-Oxo-morpholin-4-ylmethyl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5- carbonitrile 37

2-(5-Methanesulfonylmethyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carbonitrile 38

2-[5-(1-Acetyl-piperidin-4-yloxy)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5- carbonitrile 39

Cyclopropanecarboxylic acid [5-(5-cyano-2,3-dihydro-benzo[1,4]dioxin-2-yl)-pyridin- 3-ylmethyl]-amide 40

4-[5-(5-Cyano-2,3-dihydro- benzo[1,4]dioxin-2-yl)-pyridin-3-yl]-benzenesulfonamide 41

2-[5-(1,1-Dioxo-1lambda6-[1,2]thiazinan-2-ylmethyl)-pyridin-3-yl]-2,3-dihydro- benzo[1,4]dioxine-5-carbonitrile 42

2-[5-(5-Cyano-2,3-dihydro- benzo[1,4]dioxin-2-yl)-pyridin-3-ylmethoxy]-N,N-dimethyl-acetamide 43

2-[5-(2-Morpholin-4-yl-2-oxo-ethyl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5- carbonitrile 44

2-(5-Methanesulfonylmethoxy-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carbonitrile 45

2-(5-ethoxy-pyridin-3-yl)-2,3-dihydro- benzo[1,4]dioxine-5-carboxylicacid amide 46

2-{5-[(R)-(Tetrahydro-furan-3-yl)oxy]- pyridin-3-yl}-2,3-dihydro-benzo[1,4]dioxine-5-carboxylic acid amide 47

2-[5-(Tetrahydro-pyran-4-yloxy)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5- carboxylic acid amide 48

2-[5-(1-isobutyryl-piperidin-4-yloxy)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine- 5-carboxylic acid amide 49

2-[5-(2,2,2-Trifluoro-1-hydroxy-ethyl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine- 5-carboxylic acid amide 50

2-[5-(4-Hydroxy-tetrahydro-pyran-4-yl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine- 5-carbonitrile 51

2-(5-Fluoro-pyridin-3-yl)-2,3-dihydro- benzo[1,4]dioxine-5-carboxylicacid amide 52

2-(7-Hydroxy-6,7-dihydro-5H-[2]pyrindin-4-yl)-2,3-dihydro-benzo[1,4]dioxine-5- carboxylic acid amide 53

N-[5-(5-Cyano-2,3-dihydro- benzo[1,4]dioxin-2-yl)-pyridin-3-ylmethyl]-2,2,2-trifluoro-acetamide 54

Ethanesulfonic acid [5-(5-cyano-2,3-dihydro-benzo[1,4]dioxin-2-yl)-pyridin-3-ylmethyl]- amide 55

2-{5-[2-((R)-3-Hydroxy-pyrrolidin-1-yl)-2-oxo-ethyl]-pyridin-3-yl}-2,3-dihydro- benzo[1,4]dioxine-5-carbonitrile56

2-[5-Fluoro-4-((S)-1-hydroxy-ethyl)-pyridin-3-yl]-2,3-dihydrobenzo[1,4]dioxine-5- carboxylic acid amide 57

2-[5-Fluoro-4-((R)-1-hydroxy-ethyl)-pyridin-3-yl]-2,3-dihydrobenzo[1,4]dioxine-5- carboxylic acid amide 58

2-[5-Fluoro-4-(1-hydroxy-1-methyl-ethyl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine- 5-carboxylic acid amide 59

2-(5-Methyl-pyridin-3-yl)-2,3-dihydro- benzo[1,4]dioxine-5-carboxylicacid amide 60

2-[5-(cyclopropyl-ethanesulfonylamino-methyl)-pyridin-3-yl]-benzo[1,4]dioxine-5- carboxylic acid amide 61

2-(5-cyano-4-methyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylic acid amide 62

2-(5-{[imino(methyl)oxo-λ⁶- sulfanyl]methyl}pyridin-3-yl)-2,3-dihydro-1,4-benzodioxine-5-carbonitrile

In one embodiment, the invention relates to a compound selected from thegroup consisting of compounds 1-62 depicted in Table 1 above or apharmaceutically acceptable salts or a stereoisomer thereof.

In another embodiment, the invention relates to compounds 1, 5, 12, 29,37, 43, 56, 61, and 62 depicted in Table 1 above or a pharmaceuticallyacceptable salt or a stereoisomer thereof.

Unless specifically indicated, throughout the specification and theappended claims, a given chemical formula or name shall encompasstautomers and all stereo, optical and geometrical isomers (e.g.enantiomers, diastereomers, E/Z isomers, etc.) and racemates thereof aswell as mixtures in different proportions of the separate enantiomers,mixtures of diastereomers, or mixtures of any of the foregoing formswhere such isomers and enantiomers exist, as well as salts, includingpharmaceutically acceptable salts thereof and solvates thereof such asfor instance hydrates including solvates of the free compounds orsolvates of a salt of the compound.

Some of the compounds of formula (I) can exist in more than onetautomeric form. The invention includes methods for using all suchtautomers.

Compounds of the invention also include their isotopically-labelledforms. An isotopically-labelled form of an active agent of a combinationof the present invention is identical to said active agent but for thefact that one or more atoms of said active agent have been replaced byan atom or atoms having an atomic mass or mass number different from theatomic mass or mass number of said atom which is usually found innature. Examples of isotopes which are readily available commerciallyand which can be incorporated into an active agent of a combination ofthe present invention in accordance with well established procedures,include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous,fluorine and chlorine, e.g., ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³¹P, ³²P,³⁵S, ¹⁸F, and ³⁶Cl, respectively. An active agent of a combination ofthe present invention, a prodrug thereof, or a pharmaceuticallyacceptable salt of either which contains one or more of theabove-mentioned isotopes and/or other isotopes of other atoms iscontemplated to be within the scope of the present invention.

The invention includes pharmaceutically acceptable derivatives ofcompounds of formula (I). A “pharmaceutically acceptable derivative”refers to any pharmaceutically acceptable salt or ester, or any othercompound which, upon administration to a patient, is capable ofproviding (directly or indirectly) a compound useful for the invention,or a pharmacologically active metabolite or pharmacologically activeresidue thereof. A pharmacologically active metabolite shall beunderstood to mean any compound of the invention capable of beingmetabolized enzymatically or chemically. This includes, for example,hydroxylated or oxidized derivative compounds of the formula (I).

As used herein, “pharmaceutically acceptable salts” refer to derivativesof the disclosed compounds wherein the parent compound is modified bymaking acid or base salts thereof. Examples of pharmaceuticallyacceptable salts include, but are not limited to, mineral or organicacid salts of basic residues such as amines; alkali or organic salts ofacidic residues such as carboxylic acids; and the like. For example,such salts include acetates, ascorbates, benzenesulfonates, benzoates,besylates, bicarbonates, bitartrates, bromides/hydrobromides, edetates,camsylates, carbonates, chlorides/hydrochlorides, citrates, edisylates,ethane disulfonates, estolates, esylates, fumarates, gluceptates,gluconates, glutamates, glycolates, glycollylarsnilates,hexylresorcinates, hydrabamines, hydroxymaleates, hydroxynaphthoates,iodides, isothionates, lactates, lactobionates, malates, maleates,mandelates, methanesulfonates, methylbromides, methylnitrates,methylsulfates, mucates, napsylates, nitrates, oxalates, pamoates,pantothenates, phenylacetates, phosphates/diphosphates,polygalacturonates, propionates, salicylates, stearates, subacetates,succinates, sulfamides, sulfates, tannates, tartrates, teoclates,toluenesulfonates, triethiodides, ammonium, benzathines,chloroprocaines, cholines, diethanolamines, ethylenediamines, megluminesand procaines. Further pharmaceutically acceptable salts can be formedwith cations from metals like aluminium, calcium, lithium, magnesium,potassium, sodium, zinc and the like. (also see Pharmaceutical salts,Birge, S. M. et al., J. Pharm. Sci., (1977), 66, 1-19).

The pharmaceutically acceptable salts of the present invention can besynthesized from the parent compound which contains a basic or acidicmoiety by conventional chemical methods. Generally, such salts can beprepared by reacting the free acid or base forms of these compounds witha sufficient amount of the appropriate base or acid in water or in anorganic diluent like ether, ethyl acetate, ethanol, isopropanol, oracetonitrile, or a mixture thereof.

Salts of other acids than those mentioned above which for example areuseful for purifying or isolating the compounds of the present invention(e.g. trifluoro acetate salts) also comprise a part of the invention.

In addition, within the scope of the invention is use of prodrugs ofcompounds of the formula (I). Prodrugs include those compounds that,upon simple chemical transformation, are modified to produce compoundsof the invention. Simple chemical transformations include hydrolysis,oxidation and reduction. Specifically, when a prodrug is administered toa patient, the prodrug may be transformed into a compound disclosedhereinabove, thereby imparting the desired pharmacological effect.

The compounds of the invention are only those which are contemplated tobe ‘chemically stable’ as will be appreciated by those skilled in theart. For example, peroxides or a compound which would have a ‘danglingvalency’, or a ‘carbanion’ are not compounds contemplated by theinventive methods disclosed herein.

For all compounds disclosed hereinabove in this application, in theevent the nomenclature is in conflict with the structure, it shall beunderstood that the compound is defined by the structure.

All terms as used herein in this specification, unless otherwise stated,shall be understood in their ordinary meaning as known in the art. Forexample, “C₁₋₄alkyl” is a saturated aliphatic hydrocarbon monovalentradical containing 1-4 carbons such as methyl, ethyl, n-propyl,1-methylethyl (isopropyl), n-butyl or t-butyl; “C₁₋₄ alkoxy” is a C₁₋₄alkyl with a terminal oxygen, such as methoxy, ethoxy, propoxy, butoxy.All alkyl, alkenyl and alkynyl groups shall be understood as beingbranched or unbranched, cyclized or uncyclized where structurallypossible and unless otherwise specified. Other more specific definitionsare as follows:

The term “C_(1-n)-alkyl”, wherein n is an integer from 2 to n, eitheralone or in combination with another radical denotes an acyclic,saturated, branched or linear hydrocarbon radical with 1 to n C atoms.For example the term C₁₋₅-alkyl embraces the radicals H₃C—, H₃C—CH₂—,H₃C—CH₂—CH₂—, H₃C—CH(CH₃)—, H₃C—CH₂—CH₂—CH₂—, H₃C—CH₂—CH(CH₃)—,H₃C—CH(CH₃)—CH₂—, H₃C—C(CH₃)₂—, H₃C—CH₂—CH₂—CH₂—CH₂—,H₃C—CH₂—CH₂—CH(CH₃)—, H₃C—CH₂—CH(CH₃)—CH₂—, H₃C—CH(CH₃)—CH₂—CH₂—,H₃C—CH₂—C(CH₃)₂—, H₃C—C(CH₃)₂—CH₂—, H₃C—CH(CH₃)—CH(CH₃)— andH₃C—CH₂—CH(CH₂CH₃)—.

The term “C_(1-n)-alkylene” wherein n is an integer 1 to n, either aloneor in combination with another radical, denotes an acyclic, straight orbranched chain divalent alkyl radical containing from 1 to n carbonatoms. For example the term C₁₋₄-alkylene includes —(CH₂)—, —(CH₂—CH₂)—,—(CH(CH₃))—, —(CH₂—CH₂—CH₂)—, —(C(CH₃)₂)—, —(CH(CH₂CH₃))—,—(CH(CH₃)—CH₂)—, —(CH₂—CH(CH₃))—, —(CH₂—CH₂—CH₂—CH₂)—,—(CH₂—CH₂—CH(CH₃))—, —(CH(CH₃)—CH₂—CH₂)—, —(CH₂—CH(CH₃)—CH₂)—,—(CH₂—C(CH₃)₂)—, —(C(CH₃)₂—CH₂)—, —(CH(CH₃)—CH(CH₃))—,—(CH₂—CH(CH₂CH₃))—, —(CH(CH₂CH₃)—CH₂)—, —(CH(CH₂CH₂CH₃))—,—(CHCH(CH₃)₂)— and —C(CH₃)(CH₂CH₃)—.

The term “C_(3-n)-cycloalkyl”, wherein n is an integer 4 to n, eitheralone or in combination with another radical denotes a cyclic,saturated, unbranched hydrocarbon radical with 3 to n C atoms. Forexample the term C₃₋₇-cycloalkyl includes cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl and cycloheptyl.

The term “heteroatom” as used herein shall be understood to mean atomsother than carbon such as O, N, S and P.

In all alkyl groups or carbon chains one or more carbon atoms can beoptionally replaced by heteroatoms: O, S or N, it shall be understoodthat if N is not substituted then it is NH, it shall also be understoodthat the heteroatoms may replace either terminal carbon atoms orinternal carbon atoms within a branched or unbranched carbon chain. Suchgroups can be substituted as herein above described by groups such asoxo to result in definitions such as but not limited to: alkoxycarbonyl,acyl, amido and thioxo.

The term “aryl” as used herein, either alone or in combination withanother radical, denotes a carbocyclic aromatic monocyclic groupcontaining 6 carbon atoms which may be further fused to a second 5- or6-membered carbocyclic group which may be aromatic, saturated orunsaturated. Aryl includes, but is not limited to, phenyl, indanyl,indenyl, naphthyl, anthracenyl, phenanthrenyl, tetrahydronaphthyl anddihydronaphthyl.

The term “heteroaryl” means an aromatic 5 to 6-membered monocyclicheteroaryl or an aromatic 7 to 11-membered heteroaryl bicyclic ringwhere at least one of the rings is aromatic, wherein the heteroaryl ringcontains 1-4 heteroatoms such as N, O and S. Non-limiting examples of 5to 6-membered monocyclic heteroaryl rings include furanyl, oxazolyl,isoxazolyl, oxadiazolyl, thiazolyl, pyrazolyl, pyrrolyl, imidazolyl,tetrazolyl, triazolyl, thienyl, thiadiazolyl, pyridinyl, pyrimidinyl,pyridazinyl, pyrazinyl, triazinyl, and purinyl. Non-limiting examples of7 to 11-membered heteroaryl bicyclic heteroaryl rings includebenzimidazolyl, quinolinyl, dihydro-2H-quinolinyl, tetrahydroquinolinyl,isoquinolinyl, quinazolinyl, indazolyl, thieno[2,3-d]pyrimidinyl,indolyl, isoindolyl, benzofuranyl, dihydrobenzofuranyl, benzopyranyl,benzodioxolyl, benzoxazolyl and benzothiazolyl.

The term “heterocyclyl” means a stable nonaromatic 4-8 memberedmonocyclic heterocyclic radical or a stable nonaromatic 6 to 11-memberedfused bicyclic, bridged bicyclic or spirocyclic heterocyclic radical.The 5 to 11-membered heterocycle consists of carbon atoms and one ormore, preferably from one to four heteroatoms chosen from nitrogen,oxygen and sulfur. The heterocycle may be either saturated or partiallyunsaturated. Non-limiting examples of nonaromatic 4-8 memberedmonocyclic heterocyclic radicals include tetrahydrofuranyl, azetidinyl,pyrrolidinyl, pyranyl, tetrahydropyranyl, dioxanyl, thiomorpholinyl,1,1-dioxo-1λ⁶-thiomorpholinyl, morpholinyl, piperidinyl, piperazinyl,and azepinyl. Non-limiting examples of nonaromatic 6 to 11-memberedfused bicyclic radicals include octahydroindolyl, octahydrobenzofuranyl,and octahydrobenzothiophenyl. Non-limiting examples of nonaromatic 6 to11-membered bridged bicyclic radicals include2-azabicyclo[2.2.1]heptanyl, 3-azabicyclo[3.1.0]hexanyl, and3-azabicyclo[3.2.1]octanyl. Non-limiting examples of nonaromatic 6 to11-membered spirocyclic heterocyclic radicals include7-aza-spiro[3,3]heptanyl, 7-spiro[3,4]octanyl, and7-aza-spiro[3,4]octanyl. The term “heterocyclyl” or is intended toinclude all the possible isomeric forms.

The term “halogen” as used in the present specification shall beunderstood to mean bromine, chlorine, fluorine or iodine. Thedefinitions “halogenated”, “partially or fully halogenated”; partiallyor fully fluorinated; “substituted by one or more halogen atoms”,includes for example, mono, di or tri halo derivatives on one or morecarbon atoms. For alkyl, a non-limiting example would be —CH₂CHF₂, —CF₃etc.

Each alkyl, cycloalkyl, heterocycle, aryl or heteroaryl, or the analogsthereof, described herein shall be understood to be optionally partiallyor fully halogenated.

As used herein, “nitrogen” or N and “sulfur” or S includes any oxidizedform of nitrogen and sulfur and the quaternized form of any basicnitrogen. For example, for an —S—C₁₋₆ alkyl radical, unless otherwisespecified, this shall be understood to include —S(O)—C₁₋₆ alkyl and—S(O)₂—C₁₋₆ alkyl, likewise, —S—R_(a) may be represented asphenyl-S(O)_(m)— when R_(a) is phenyl and where m is 0, 1 or 2.

General Synthetic Methods

The compounds of the invention may be prepared by the methods andexamples presented below and methods known to those of ordinary skill inthe art. The methods that are described here are intended as anillustration and for the enablement of the instant invention withoutrestricting the scope of its subject matter, the claimed compounds, andthe examples. Optimum reaction conditions and reaction times may varydepending on the particular reactants used. Unless otherwise specified,solvents, temperatures, pressures, and other reaction conditions may bereadily selected by one of ordinary skill in the art. Specificprocedures are provided below. Intermediates used in the syntheses beloware either commercially available or easily prepared by methods known tothose skilled in the art. Reaction progress may be monitored byconventional methods such as thin layer chromatography (TLC) or highpressure liquid chromatography-mass spec (HPLC-MS). Intermediates andproducts may be purified by methods known in the art, including columnchromatography, HPLC, preparative TLC, supercritical fluidchromatography (SFC), and recrystallization.

Compounds of formula (I) may be prepared as illustrated in Scheme 1.

As illustrated in Scheme 1, a suitable heteroaromatic bromide may beconverted to boronate ester II via palladium catalyzed coupling reactionwith a diboronyl ester such as bis(pinacolato)diboron. Suzuki reactionwith vinyl bromide III (Intermediate 1) provides IV. Aminolysis of esterIV provides amide V. Hydrogenation over palladium on carbon provides thedesired compound of formula I (R¹═CONH₂).

Compounds of formula (I) may also be prepared as illustrated in Scheme2.

As illustrated in Scheme 2, compounds of formula I may also be preparedby hydrogenation of compound IV followed by aminolysis to give I.

Compounds of formula (I) may also be prepared as illustrated in Scheme3.

As illustrated in Scheme 3, a suitable heteroaromatic bromide may beconverted to a boronate ester II via palladium catalyzed couplingreaction with a diboronyl ester such as bis(pinacolato)diboron. Suzukireaction with vinyl bromide VII (Intermediate 2) provides VIII.Hydrogenation over palladium on carbon provides the desired compound offormula I (R¹═CONH₂).

Compounds of formula I R¹═—CN maybe prepared from compounds of formula 1R¹═—CONH₂ by reacting with a suitable dehydrating reagent such astrifluoroactetic anhydride in the presence of base as shown is Scheme 4.

SYNTHETIC EXAMPLES Synthesis of Intermediates Intermediate 1:2-bromo-benzo[1,4]dioxine-5-carboxylic acid methyl ester

Step A: To a suspension of 2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid (49.7 g, 275.6 mmol) in 1000 mL of MeOH, is added acetyl chloride(40.0 ml, 560.5 mmol) in a drop-wise manner. Upon complete addition, thereaction is stirred at room temperature for 18 hours. The reactionmixture is then concentrated in vacuo and the residue is dissolved inEtOAc and washed with sat. NaHCO₃. The aqueous layer is separated, andextracted with EtOAc. The combined organic layers are washed with brine,dried under Na₂SO₄, filtered and concentrated to afford 50.7 g of2,3-dihydro-benzo[1,4]dioxine-5-carboxylic acid methyl ester.

Step B: To a mixture of 2,3-dihydro-benzo[1,4]dioxine-5-carboxylic acidmethyl ester (50.7 g, 261.1 mmol) in carbon tetrachloride (300 ml) isadded 2,2′-azobis(isobutryonitrile) (125 mg, 0.7 mmol) andN-bromosuccinimide (90.0 g, 505.7 mmol). The reaction mixture isrefluxed using a 60 W lamp (covered with aluminum foil) for 24 hours.After this time another 100.0 g (561.8 mmol) of N-bromosuccinimide, 175mg (1.1 mmol) of 2,2′-azobis(isobutryonitrile) and 100 mL of carbontetrachloride are added. The reaction mixture is stirred under the sameconditions for another 24 hours. After this time, another 40.0 g (224.7mmol) of N-bromosuccinimide and 100 mg (0.6 mmol) of2,2′-azobis(isobutryonitrile) are added. The reaction mixture is stirredunder the same conditions for another 72 hours after which time thereaction appeared complete. To the reaction mixture is added 1 L ofether. The resulting solid is filtered off and washed with ether. Thecombined organics are concentrated and the crude solid is dissolved in20% EtOAc/heptane, and purified by a plug of silica gel (500 g), elutingwith 20% EtOAc/heptane. The product fractions are collected andconcentrated to afford 86.5 g of2,3-dibromo-2,3-dihydro-benzo[1,4]dioxine-5-carboxylic acid methylester.

Step C: A suspension of2,3-dibromo-2,3-dihydro-benzo[1,4]dioxine-5-carboxylic acid methyl ester(22.7 g, 64.5 mmol) in 200 mL of MeOH is warmed to 50° C. and treatedwith 500 mL of sodium methoxide (0.5M in methanol, 250 mmol). Thereaction mixture is warmed to 65° C. and stirred for 2 hours. Thereaction mixture is treated with silica gel and concentrated. The dryresidue is purified via silica gel flash column chromatography elutingwith 0-15% EtOAc/heptane to afford 2.6 g of the title compound.

Intermediate 2: 2-bromo-benzo[1,4]dioxine-5-carboxylic acid amide

A 20 mL reaction vessel is charged with2-bromo-benzo[1,4]dioxine-5-carboxylic acid methyl ester (1.2 g, 4.4mmol) and 7N ammonia solution in methanol (13.0 mL, 88.5 mmol). Thevessel is capped and heated at 75 C for 18 hours. Upon cooling to roomtemperature, the mixture is concentrated to dryness. The remaining solidis diluted with MeOH (10 mL) and sonicated. Filtration affords 1.00 g of2-bromo-benzo[1,4]dioxine-5-carboxylic acid amide.

Intermediate 3: 2-bromo-benzo[1,4]dioxine-5-carboxylic acid methylamide

The title compound is prepared in a similar manner to Intermediate 2replacing ammonia with methylamine.

Intermediate 4: 3-bromo-5-fluoro-4-methyl-pyridine

A solution of diisopropylamine (1.9 mL, 13.7 mmol) in 20 mL of THF iscooled to 0° C. and treated with n-butyllithium (6.7 mL, 13.6 mmol). Themixture is stirred at 0° C. for 15 minutes then cooled to −78° C.3-Bromo-5-fluoropyridine (2.0 g, 11.4 mmol) is added drop-wise as asolution in 20 mL of THF. This mixture is stirred at −78° C. for 45minutes. A separate solution of iodomethane (2.1 mL, 34.1 mmol) in 20 mLof THF is cooled to −78° C. The anion solution is then cannulated intothe iodomethane solution. Once the transfer is complete, the mixture isstirred at −78° C. for 30 minutes. The cooling bath is removed and themixture is stirred for 30 minutes and then quenched with saturated NH₄Clsolution. The mixture is diluted with EtOAc and water. The organic layeris washed with brine, dried over MgSO₄, filtered and concentrated. Theresidue is purified via silica gel flash column chromatography elutingwith 0-10% EtOAc/heptane to afford 1.4 g of the title compound.

Intermediate 5: 3-bromo-5-difluoromethyl-pyridine

A solution of 5-bromo-3-formylpyridine (1.5 g, 8.1 mmol) in 15.00 mL ofDCM is cooled to −78° C. and then treated with diethylaminosulfurtrifluoride (5.3 mL, 40.3 mmol) drop-wise. The solution is allowed towarm to room temperature overnight. The reaction mixture is addeddrop-wise to a stirred cold solution of dilute NH₄OH and diluted withmore DCM. The organic layer is separate and the aqueous layer is backextracted with DCM. The organic layers are combined and are washed withbrine, dried over MgSO4, filtered and concentrated. The residue ispurified by silica gel flash column chromatography eluting with 0-30%EtOAc/heptane to afford 1.1 g of the title compound.

Intermediate 6: 3-[(5-bromo-3-pyridyl)methyl]oxazolidin-2-one

Step A: A solution of (5-bromo-3-pyridyl)methanol (7.0 g, 37.2 mmol) in10 mL of DCM is cooled to 0° C. Triphenylphosphine (9.8 g, 37.2 mmol) isadded followed by the slow addition of carbon tetrabromide (18.5 g, 55.8mmol) as the reaction is exothermic. The mixture is stirred at 0′C for 3hours. After the reaction is complete, the reaction mixture is absorbedwith silica gel and purified by silica flash column chromatography toafford 7.5 g of 3-bromo-5-(bromomethyl)pyridine.

Step B: 2-Oxazolidone (0.6 g, 7.2 mmol) is dissolved in 20 mL of DMF andcooled to 0° C. 60% sodium hydride (0.29 g, 7.2 mmol) is added. Bubblingis observed. The mixture is stirred for 5 minutes.3-Bromo-5-(bromomethyl)pyridine (1.2 g, 4.8 mmol) as a solution in 15 mLof DMF is added slowly. The reaction mixture is allowed to warm to roomtemperature for 16 hours. The reaction is quenched with 10 mL of water.The mixture is filtered through diatomaceous earth and rinsed with EtOAc(50 mL). The EtOAc layer is concentrated. The crude product is by silicagel flash column chromatography eluting with 0-10% MeOH in DCM to afford0.9 g of the title compound.

The following intermediates are synthesized according to the procedurefor Intermediate 6, substituting the appropriate commercially availablereagents.

Intermediate Structure Name 7

1-(5-bromo-pyridin-3-ylmethyl)-pyrrolidin-2-one 8

4-[5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-3-ylmethyl]-morpholin-3-one 9

2-(5-bromo-pyridin-3-ylmethyl)-[1,2]thiazinane 1,1-dioxide

Intermediate 10: 4-(5-bromo-pyridin-3-yl)-benzenesulfonamide

3,5-Dibromopyridine (1.0 g, 4.2 mmol), (4-aminosulphonyl)benzeneboronicacid (0.8 g, 4.2 mmol),1,1′-bis(diphenylphosphino)ferrocenedichloropalladium(II) DCM complex(172 mg, 0.211 mmol), 20 mL of 1,4-dioxane, and 2.0M sodium carbonatesolution (4.2 mL, 8.4 mmol) are combined in a pressure vessel. Thevessel is flushed with argon, sealed and stirred at 120° C. for 2 hours.The reaction mixture is diluted with EtOAc/water. The mixture isfiltered through diatomaceous earth, and the layers are separated. Theorganic layer is washed with brine, dried over MgSO₄, filtered andconcentrated. The residue is purified by silica gel flash columnchromatography eluting with 50-100% EtOAc/Heptane to afford 0.6 g of thetitle compound.

Intermediate 11:1-[(S)-3-(5-bromo-pyridin-3-yloxy)-pyrrolidin-1-yl]-ethanone

To a stirred solution of triphenylphosphine (28.9 g, 110 mmol) in 100 mLof THF cooled to 0° C. is added diisopropyl azodicarboxylate (20.9 g,103 mmol) and 5-bromo-pyridin-3-ol (12.0 g, 69 mmol) as a solution in 50mL THF. 1-((R)-3-Hydroxy-cyclopentyl)-ethanone (8.8 g, 69 mmol) as asolution in 50 mL of THF is added slowly. The reaction is stirred atroom temperature for 3 hours. The reaction is quenched with water andextracted with EtOAc (2×200 mL). The combined organic layers areconcentrated under reduced pressure. The crude product is purified bysilica gel flash chromatography and washed with diethyl ether to afford6.5 g of the title compound.

The following intermediates are synthesized according to the procedurefor Intermediate 10, substituting the appropriate commercially availablereagents.

Intermediate Structure Name 12

1-[(R)-3-(5-bromo-pyridin-3-yloxy)-pyrrolidin-1- yl]-ethanone 13

1-[3-(5-bromo-pyridin-3-yloxy)-azetidin-1-yl]- ethanone 14

1-[4-(5-bromo-pyridin-3-yloxy)-piperidin-1-yl]- ethanone

Intermediate 15: 3-bromo-5-methanesulfonylmethyl-pyridine

Step A: To a cooled (0° C.) solution of (5-bromo-pyridin-3-yl)-methanol(5.0 g, 26.6 mmol) and triphenylphosphine (8.4 g, 31.9 mmol) in 130 mLof DCM is added carbon tetrabromide (13.2 g, 39.9 mmol). The resultingmixture is stirred at 0° C. for 10 minutes. The mixture is concentratedand purified by silica gel flash chromatography eluting with 0-40% EtOAcin heptane to afford 6.1 g of 3-bromo-5-bromomethyl-pyridine.

Step B: 3-Bromo-5-bromomethyl-pyridine (100 mg, 0.4 mmol), sodiummethanesulphinate (122 mg, 1.2 mmol), and 1 mL of DMF are combined in areaction vial. The vial is sealed and the reaction is stirred at 65° C.in a heating block for 1 hour. The mixture is cooled to roomtemperature, diluted with EtOAc (30 mL), washed with water (3×15 mL),and brine, dried over sodium sulfate, filtered, and concentrated. Thecrude product is purified by silica gel flash chromatography elutingwith 0-100% EtOAc in heptane to afford 70 mg of the title compound.

Intermediate 16:1-[(R)-3-(5-bromo-pyridin-3-yloxy)-piperidin-1-yl]-ethanone

Step A: To a cooled (0° C.) solution of PPh₃ (1.2 g, 4.5 mmol) in 50 mLof THF is added diisopropyl azodicarboxylate (0.81 mL, 4.1 mmol),drop-wise. After stirring at 0° C. for 15 minutes, 5-bromo-pyridin-3-ol(441 mg, 2.5 mmol) and (S)-3-hydroxy-piperidine-1-carboxylic acidtert-butyl ester (500 mg, 2.5 mmol) are added and the mixture is warmedand stirred at room temperature for 16 hours. The mixture isconcentrated and purified by silica gel flash column chromatography togive 596 mg of (R)-3-(5-bromo-pyridin-3-yloxy)-piperidine-1-carboxylicacid tert-butyl ester.

Step B: A solution of(R)-3-(5-bromo-pyridin-3-yloxy)-piperidine-1-carboxylic acid tert-butylester (596 mg, 1.7 mmol) in 5 mL of MeOH and 4 N HCl solution in1,4-dioxane (1.5 mL) is stirred at room temperature for 16 hours. Themixture is concentrated to provide 525 mg of3-bromo-5-((R)-piperidin-3-yloxy)-pyridine as the hydrochloride salt.

Step C: To a solution of 3-bromo-5-((R)-piperidin-3-yloxy)-pyridinehydrochloride salt (525 mg, 1.8 mmol) in 10 mL of DMF is added acetylchloride (0.19 mL, 2.7 mmol) and N,N-diisopropylethylamine (1.4 mL, 8.0mmol). The mixture is stirred at room temperature for 16 hours. Thereaction is partitioned between H₂O and EtOAc, and the layers areseparated. The aqueous layer is extracted with EtOAc. The organic layersare combined, dried and concentrated. The crude product is purified bysilica gel flash column chromatography to provide 271 mg of the titlecompound.

Intermediate 17: methanesulfonic acid5-(tetrahydro-pyran-4-yl)-pyridin-3-yl ester

Step A: 5-Bromo-pyridin-3-ol (15 g, 86.2 mmol),4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-3,6-dihydro-2H-pyran(27 g, 129.3 mmol), potassium acetate (12.7 g, 129.3 mmol), andBis(diphenylphosphino)ferrocene]dichloropalladium(II) (1.3 g, 1.7 mmol)are combined in 150 mL of dioxane and 30 mL of water. The reaction isrefluxed for 16 hours. The reaction is concentrated to dryness. Theresidue is partitioned between H₂O and EtOAc and the layers areseparated. The aqueous layer is extracted with EtOAc and the combinedorganic layers are dried and concentrated. The crude product is purifiedby silica gel flash column chromatography to provide 10.5 grams of5-(3,6-dihydro-2H-pyran-4-yl)-pyridin-3-ol.

Step B: To the solution of 5-(3,6-dihydro-2H-pyran-4-yl)-pyridin-3-ol(9.0 g, 50.8 mmol) in one liter of MeOH is added 10% Pd—C. Thesuspension is degassed under vacuum and is purged with hydrogen. Themixture is stirred under 50 psi of hydrogen at 50° C. for 5 hours. Atthe end of the reaction, the mixture is filtered and washed with MeOH.The filtrate is concentrated and purified by silica gel flash columnchromatography to give 9 grams of5-(tetrahydro-pyran-4-yl)-pyridin-3-ol.

Step C: To a solution of the 5-(tetrahydro-pyran-4-yl)-pyridin-3-ol (500mg, 2.8 mmol), DMAP (13 mg, 0.1 mmol), and triethylamine (0.78 mL, 5.6mmol) in 20 mL of DCM is added triflic anhydride (0.47 mL, 2.8 mmol)drop-wise. The reaction is allowed to stir at room temperatureovernight. The reaction is diluted with 1N NaOH. The layers areseparated and the DCM layer is concentrated to dryness. The residue ispurified by silica gel flash column chromatography eluting with 5-50%EtOAc in heptanes to give 415 mg of the title compound.

Intermediate 18: 1-(5-bromo-pyridin-3-yl)-cyclohexanol

To 3,5-dibromopyridine (1.5 g, 6.3 mmol) in 6 mL of THF at −20° C. isadded 1.3M i-PrMgCl_LiCl solution (4.7 mL, 6.1 mmol) in one portion. Themixture is allowed to stir for 30 minutes, warming to −10° C. Themixture is cooled to −20° C. and cyclohexanone (0.79 mL, 7.6 mmol) isadded. The reaction is quenched with 50 mL of saturated aqueous NH₄Cland diluted with 200 mL EtOAc. The organic phase is washed with 2×100 mLof H₂O and 1×100 mL of brine. The organic phase is dried with MgSO₄,filtered and concentrated. The residue is purified by silica gel flashcolumn chromatography eluting with 0-10% MeOH/CH₂Cl₂ to give 560 mg ofthe title compound.

The following intermediates are synthesized according to the procedurefor Intermediate 18, substituting either commercially available reagentsor the appropriate intermediates described above.

Intermediate Structure Name 19

1-(5-bromo-pyridin-3-yl)-cyclobutanol 20

4-(5-bromo-pyridin-3-yl)-tetrahydropyran-ol

Intermediate 21: 5-(5-bromo-pyridin-3-yl)-1-methyl-pyrrolidin-2-one

Step A: 3-Bromo-5-(pyrrolidin-2-yl)pyridine (400 mg, 1.8 mmol), 4 mL ofglacial acetic acid and 1 mL of water is added to a reaction vial.Bromine (0.8 mL) is added drop-wise. The vial is sealed and the reactionis heated to 90° C. in an oil bath and continued to stir at thattemperature for 3 hours. The mixture is cooled to room temperature.Water (15 mL) is added to the cooled reaction mixture and the mixture issaturated with solid potassium carbonate. The mixture is extracted withEtOAc (3×30 mL). The combined organics are dried over sodium sulfate,filtered, and concentrated. The residue is purified by silica gel flashcolumn chromatography eluting with 0-6% MeOH in DCM to afford 0.65 g of3,3-dibromo-5-(5-bromo-pyridin-3-yl)-pyrrolidin-2-one.

Step B: Sodium borohydride (0.74 g, 19.6 mmol) is suspended in 17 mL ofethanol and tellurium metal powder (1.25 g, 9.8 mmol) is added inportions. The mixture is heated under reflux for 15 minutes and themixture becomes a light purple color. The mixture is cooled to roomtemperature. 3,3-Dibromo-5-(5-bromo-pyridin-3-yl)-pyrrolidin-2-one (0.65g, 1.6 mmol) dissolved in 5 mL of ethanol is added slowly. The mixtureis stirred at room temperature for 72 hours. The mixture is filteredthrough diatomaceous earth and washed with MeOH. The filtrate isconcentrated. The resulting crude product is purified by silica gelflash column chromatography eluting with 0-6% MeOH in DCM to afford 290mg of 5-(5-bromo-pyridin-3-yl)-pyrrolidin-2-one.

Step C: To a solution of 5-(5-bromo-pyridin-3-yl)-pyrrolidin-2-one (202mg, 0.84 mmol) in 5 mL of THF is added 60% NaH (50 mg, 1.3 mmol). Themixture is stirred at room temperature for 5 minutes and methyl iodide(0.078 mL, 1.3 mmol) is then added drop-wise. The mixture is stirred atroom temperature for 16 hours. The mixture is then concentrated andpurified by silica gel flash column chromatography to give 157 mg of thetitle compound. Enantiomers are separated using Chiral SFC (ChiralpakAD-H, 30% (1:1 Isopropanol+0.5% TFA:Hexanes):CO₂, 70 mL/min, 140 bar,25° C.).

Intermediate 22: 1-(5-Bromo-4-methyl-pyridin-3-yl)-ethanol

To a solution of 3,5-dibromo-4-methyl-pyridine (2.0 g, 8.0 mmol) in 100mL of THF cooled in a liquid N₂/ethanol bath below −100° C. is added 2.5M n-butyllithium in hexanes solution (3.2 mL, 8.0 mmol). This is stirredfor 5 minutes, then neat acetaldehyde (4.5 mL, 8.0 mmol) is added all atonce. The reaction is allowed to warm to −78° C. over 30 minutes. Thetemperature is held at −78° C. by adding dry ice to the bath. Thereaction is kept at −78° C. for 1 hour. The reaction is quenched withsat NH₄Cl at −78° C. The reaction is allowed to warm to roomtemperature. The reaction is diluted with EtOAc and water. The organiclayer is concentrated to dryness. The residue is purified by silica gelflash column chromatography eluting with 20-100% EtOAc in heptanes togive 0.88 g of the title compound. Chiral SFC (LUX Cellulose-4,12%(1:1:1 MeOH:EtOH:IPA):CO₂, 70 mL/min, 120 bar, 40° C.) of 2.5 g of1-(5-bromo-4-methyl-pyridin-3-yl)-ethanol gives 0.98 g of enantiomer Aand 0.98 g of enantiomer B.

Intermediate 23: 3-bromo-5-methanesulfonylmethoxy-pyridine

Step A: To a solution of 5-bromo-pyridin-3-ol (500 mg, 2.9 mmol) in 5 mLof DMF is added sodium hydride 60% dispersion in mineral oil (230 mg,5.8 mmol). The reaction is stirred for 15 minutes when chloromethylmethyl sulfide (0.24 mL, 2.9 mmol) is added. The reaction is stirred for1 hour, then it is diluted with EtOAc and water. The organic layer isconcentrated to dryness to give 330 mg of3-bromo-5-methylsulfanylmethoxy-pyridine.

Step B: To a solution of 3-bromo-5-methylsulfanylmethoxy-pyridine (330mg, 1.4 mmol) in 10 mL of DCM is added 3-chloroperbenzoic acid 77% (608mg, 3.5 mmol). The reaction is allowed to stir overnight. The mixture isquenched with 1N NaOH. The layers are separated and the organic layer isconcentrated to dryness. Silica gel flash column chromatography elutingwith EtOAc in heptanes gives 175 mg of the title compound.

Intermediate 24: 2-[(5-bromo-3-pyridyl)methoxy]-N,N-dimethyl-acetamide

(5-Bromo-pyridin-3-yl)-methanol (2.0 g, 11 mmol) is added to a 0° C.solution of 60% NaH (0.51 g, 12.8 mmol) in 150 mL of THF. The mixture isstirred at room temperature for 1 hour then cooled to 0° C.2-Chloro-N,N-dimethyl-acetamide (1.42 g, 12 mmol) is added to themixture. The cooling bath is removed and the mixture is stirred at roomtemperature for 16 hours. The reaction is quenched with brine (0.5 mL)and filtered through a pad of diatomaceous earth. The filtrate isconcentrated, diluted with DCM, treated with MgSO₄ and filtered throughdiatomaceous earth again. The filtrate is concentrated and the crudeproduct is purified by silica gel flash column chromatography elutingwith 0-6% MeOH/DCM to 1.95 g of the title compound.

Intermediate 25: 2-(5-bromo-3-pyridyl)-1-morpholino-ethanone

To the solution of 5-bromo-3-pyridineacetic acid (500 mg, 2.3 mmol) in 3mL of DMF is added TBTU (1.1 g, 3.4 mmol). Morpholine (0.61 mL, 6.9mmol) is added drop-wise. The resulting reaction mixture is stirred atroom temperature for 16 hours. The mixture is diluted with 50 mL ofEtOAc, washed with water (3×5 mL), and brine, dried over sodium sulfate,filtered, and concentrated. The resulting crude product is purified bysilica gel flash column chromatography eluting with 0-4.5% MeOH/DCM toafford 381 mg of the title compound.

The following intermediates are synthesized according to the procedurefor Intermediate 25, substituting either commercially available reagentsor the appropriate intermediates described above.

Intermediate Structure Name 26

2-(5-bromo-3-pyridyl)-1-(4,4-difluoro-1- piperidyl)ethanone 27

2-(5-bromo-pyridin-3-yl)-N,N-dimethyl- acetamide 28

2-(5-bromo-pyridin-3-yl)-1-(R)-3-hydroxy- pyrrolidin-1-yl)-ethanone

Intermediate 29: 1-(5-bromo-pyridin-3-yl)-cyclopropanecarbonitrile

To a suspension of (5-bromo-pyridin-3-yl)-acetonitrile (1.0 g, 5.1 mmol)in 50% NaOH (20 mL) is added 1-bromo-2-chloro-ethane (764 mg, 5.3 mmol)and benzyl triethylammonium chloride (15 mg, 0.1 mmol). The resultantmixture is heated to 60° C. for 2 hours. After cooling down to roomtemperature, EtOAc is added. The layers are separated, and the aqueouslayer is extracted with fresh EtOAc. The organic layers are combined,washed with brine, dried over Na₂SO₄, filtered and concentrated. Theproduct is purified by silica gel flash column chromatography to afford626 mg of the title compound.

Intermediate 30: cyclopropanecarboxylic acid(5-bromo-pyridin-3-ylmethyl)-amide

To a stirred solution of cyclopropanecarboxylic acid (0.58 g, 6.7 mmol)in 50 mL of DMF is added HATU (3.1 g, 8.0 mmol) followed by(5-bromo-pyridin-3yl)-methylamine (1.3 g, 6.7 mmol) andN,N-diisopropylethylamine (7.5 mL, 42.8 mmol). The resulting mixture isstirred at room temperature for 16 hours after which time it isconcentrated to low volume, poured into 150 mL of water and extractedwith EtOAc (3×). The combined organics are dried over MgSO₄, filteredand concentrated. The remaining residue is purified via silica gel flashcolumn chromatography eluting with 0-8% MeOH/DCM to give 1.10 g of thetitle compound.

The following intermediate is synthesized according to the procedure forIntermediate 30, substituting the appropriate commercially availablereagent.

Intermediate Structure Name 31

cyclopropanecarboxylic acid (5-bromo-pyridin-3- ylmethyl)-methyl-amide

Intermediate 32: 3-bromo-5-(4-fluoro-tetrahydro-pyran-4-yl)-pyridine

A solution of (diethylamino)sulfur trifluoride (0.63 g, 3.9 mmol) in 6.0mL of DCM is cooled to −78° C. and treated with a solution of4-(5-bromo-pyridin-3-yl)-tetrahydro-pyran-4-ol (1.0 g, 3.9 mmol) in 15mL of DCM. The reaction is stirred at −78° C. for 2 hours then warmed toroom temperature and poured over ice. The mixture is stirred until allof the ice has melted at which time the layers are separated. Theaqueous phase is extracted once more with DCM and the combined organicsare washed with water, brine and then dried (MgSO₄). The organic isfiltered and concentrated to give 0.90 g of the title compound.

Intermediate 33: 1-(5-bromo-pyridin-3-yl)-2,2,2-trifluoro-ethanol

To a cooled (0° C.) solution of 5-bromo-pyridine-3-carboxaldehyde (2.0g, 10.8 mmol) in 25 mL of THF is added trimethyl(trifluoromethyl)silane(2.8 mL, 18.8 mmol) and 1.0M TBAF in THF solution (10.8 mL, 10.8 mmol).The mixture is warmed to room temperature for 3 hours. The solvent isevaporated to give the crude product. Purification by silica gel flashcolumn chromatography affords 1.9 g of the title compound.

Intermediate 34: 4-Bromo-6,7-dihydro-5H-[2]pyrindin-7-ol

Step A: A solution of diisopropylamine (3.37 mL, 23.9 mmol) in 100 mL ofTHF is cooled to 0° C. and then treated with n-butyllithium (11.95 mL,23.9 mmol). The mixture is stirred at 0° C. for 15 minutes then cooledto −78° C. Methyl 5-bromonicotinate (4.70 g, 21.7 mmol) is added assolution in 20 mL of THF drop-wise. The mixture is stirred at −78° C.for 30 minutes then treated with methyl acrylate (4.89 mL, 54.3 mmol) in20 mL of THF drop-wise. The mixture is stirred at −78° C. for 1.5 hoursthen quenched with 50 mL of 10% acetic acid. The reaction mixture isevaporated to dryness. The crude solid is treated with 54 mL of 6N HCland stirred at 100° C. for 1 hour. The reaction mixture is cooled inice, basified to pH 7-8 with 5N NaOH and extracted twice with EtOAc. Thecombined org layer is washed with brine, dried over MgSO₄, filtered andconcentrated. Purification by silica gel flash column chromatographyeluting with 20-50% EtOAc/heptane affords 817 mg of4-bromo-5,6-dihydro-[2]pyrindin-7-one.

Step B: A mixture of 4-bromo-5,6-dihydro-[2]pyrindin-7-one (1.96 g, 9.2mmol) in 100 mL of ethanol is cooled to 0° C. and then treated withsodium borohydride (454.58 mg, 12.0 mmol). The reaction is stirred atroom temperature for 1 hour and the solvent is evaporated. The crudesolid is taken into EtOAc/water and the layers are separated. The orglayer is washed with brine, dried over MgSO₄, filtered and concentrated.Purification by silica gel flash column chromatography eluting with50-100% EtOAc/heptane affords 1.5 g of the title compound.

Synthesis of Final Compounds

Chiral SFC conditions for enantiomer resolution are set forth in Table2. When absolute stereochemistry is not established, by definition, thefirst-eluting enantiomer is referred to as enantiomer A, and thesecond-eluting enantiomer is referred to as enantiomer B. Where acompound contains two stereocenters, the diastereomers are designatedAA, AB, BA, and BB, with the first letter referring to the firstresolved stereocenter and the second letter referring to the secondresolved stereocenter in a given synthetic sequence, with A and Bdesignations for order of elution as above. LCMS data are measured usingthe methods set forth in Table 3. LCMS Data for the compounds in Table 1are shown in Table 4. Compounds that were separated into theirenantiomers are shown by separate entries in Tables 4 and 5 forenantiomer A and enantiomer B Likewise, compounds that were separatedinto their diastereomers are shown by separate entries for diastereomersAA, AB, BA, and BB.

Example 1:2-[5-(4-Hydroxy-tetrahydro-pyran-4-yl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid amide. (Cpd 1, Table 1)

Step A: 4-(5-Bromo-pyridin-3-yl)-tetrahydro-pyran-4-ol (516 mg, 2.0mmol), bis(pinacolato)diboron (760 mg, 3.0 mmol), potassium acetate (785mg, 8.0 mmol), and Bis(diphenylphosphino)ferrocene]dichloropalladium(II)(146 mg, 0.2 mmol) are combined in a vial. Dioxane (5 nit) is added andAr is bubbled through the mixture for 5 minutes. The vial is capped andheated at 80° C. for 4 hours to provide4-[5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-3-yl]-tetrahydro-pyran-4-ol.This is used in situ for the subsequent Suzuki coupling.

Step B: To the above mixture of4-[5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-3-yl]-tetrahydro-pyran-4-olis added 2-bromo-benzo[1,4]dioxine-5-carboxylic acid methyl ester (540mg, 2.0 mmol),[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(III) (146 mg,0.2 mmol), and 2M aqueous sodium carbonate solution (2.0 ml, 4.0 mmol).Ar is bubbled through the mixture for 5 minutes. The vial is capped andheated at 80° C. for 16 hours. The reaction is cooled to roomtemperature and is poured into water. This is extracted three times withEtOAc. The combined organic extracts are washed with brine, dried(Na₂SO₄), filtered, and concentrated to dryness. The crude product ispurified by silica gel flash chromatography eluting with 1-5% MeOH inDCM to provide 290 mg of2-[5-(4-hydroxy-tetrahydro-pyran-4-yl)-pyridin-3-yl]-benzo[1,4]dioxine-5-carboxylicacid methyl ester.

Step C: A mixture of2-[5-(4-hydroxy-tetrahydro-pyran-4-yl)-pyridin-3-yl]-benzo[1,4]dioxine-5-carboxylicacid methyl ester (100 mg, 0.3 mmol) and 10% palladium on carbon,Degussa type (50 mg) in 1 mL of acetic is degassed and placed under aballoon of hydrogen. The reaction is stirred at room temperature for 4hours. The catalyst is filtered off and washed with methanol. Thefiltrate is concentrated to dryness. The residue is diluted with EtOAcand washed with 1N NaOH and brine. The EtOAc layer is dried (Na₂SO₄),filtered, and concentrated to dryness. The crude product is purified viasilica gel flash column chromatography eluting with 1-5% MeOH in DCM toprovide 60 mg of2-[5-(4-hydroxy-tetrahydro-pyran-4-yl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester.

Step D: A mixture of2-[5-(4-hydroxy-tetrahydro-pyran-4-yl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester (400 mg, 1.1 mmol) and 7 N ammonia in methanolsolution (3 mL, 20 mmol) is heated in a sealed tube at 70° C. for 7days. The reaction is concentrated to dryness. The residue is purifiedby flash chromatography on a Biotage KP-NH column (1-5% MeOH in DCM) toprovide 300 mg of the title compound. The stereoisomers are separatedusing chiral SFC.

Compound 2 in Table 1 is synthesized according to the procedure outlinedin Example 1, substituting either commercially available reagents or theappropriate intermediates described above.

Example 2:2-[5-(1,1-Dioxo-1λ⁶,-[1,2]thiazinan-2-ylmethyl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid amide (Cpd 3, Table 1)

Step A: 2-(5-Bromo-pyridin-3-ylmethyl)-[1,2]thiazinane 1,1-dioxide (1.5g, 5.0 mmol), bis(pinacolato)diboron (1.90 g, 7.5 mmol), potassiumacetate (1.96 g, 20.0 mmol),[1,1′-bis(diphenylphosphino)ferrocene)dichloropalladium(II) (365.86 mg,0.5 mmol) and 16 mL of 1,4-dioxane are combined in a reaction vessel.The vessel is flushed with argon and sealed. The mixture is stirred at120° C. for 2 hours and cooled to room temperature.

Step B: 2-Bromo-benzo[1,4]dioxine-5-carboxylic acid methyl ester (1.00g, 3.7 mmol) is added to the reaction mixture from step A, followed by5.0 mL of 1,4-dioxane and 2M aqueous sodium carbonate (3.7 mL, 7.4mmol). The vessel is flushed with argon and sealed. The mixture isstirred at 100° C. for 16 hours. The reaction mixture is diluted withEtOAc and water and filtered through diatomaceous earth. The layers areseparated and the organic layer is washed with brine, dried over MgSO₄,filtered and concentrated. The crude product is purified by silica gelflash column chromatography eluting with EtOAc to give 1.1 g ofdioxo-1λ⁶,-[1,2]thiazinan-2-ylmethyl)-pyridin-3-yl]-benzo[1,4]dioxine-5-carboxylicacid methyl ester.

Step C:2-[5-(1,1-Dioxo-1λ⁶,-[1,2]thiazinan-2-ylmethyl)-pyridin-3-yl]-benzo[1,4]dioxine-5-carboxylicacid methyl ester (1.1 g, 2.6 mmol) and 7N ammonia in MeOH solution(18.6 ml, 130.3 mmol) are combined in pressure vessel. The vessel issealed and stirred at 85° C. for 16 hours. The resulting gray solid isfiltered to give 656 mg of2-[5-(1,1-dioxo-1λ⁶,-[1,2]thiazinan-2-ylmethyl)-pyridin-3-yl]-benzo[1,4]dioxine-5-carboxylicacid amide.

Step D:2-[5-(1,1-Dioxo-1λ⁶,-[1,2]thiazinan-2-ylmethyl)-pyridin-3-yl]-benzo[1,4]dioxine-5-carboxylicacid amide (630 mg, 1.6 mmol), 50 mL of acetic acid and 10% palladium oncarbon (167 mg, 0.16 mmol) are combined. The mixture is stirred under anatmosphere of hydrogen for 3 hours at room temperature and the reactionmixture is filtered through diatomaceous earth. The filtrate isconcentrated and the crude solid is purified by silica gel flash columnchromatography eluting with 50-100% EtOAc/10% MeOH/EtOAc to give 375 mgof the title compound. The stereoisomers are separated by chiral SFC.

Compounds 4 through 27 and compounds 51 and 59 in Table 1 aresynthesized according to the procedure for Example 2, substitutingeither commercially available reagents or the appropriate intermediatesdescribed above.

Compound 28 in Table 1 is synthesized according to the procedure forExample 2, substituting the appropriate commercially available reagentand 33% methylamine in ethanol for ammonia in methanol in Step C.

Example 3:2-[5-[2-(Dimethylamino)-2-oxo-ethyl]-3-pyridyl]-2,3-dihydro-1,4-benzodioxine-5-carboxamide(Cpd 29, Table 1)

Step A: 2-(5-Bromo-3-pyridyl)-N,N-dimethyl-acetamide (0.8 g, 3.3 mmol),bis(pinacolato)diboron (1.0 g, 4.1 mmol), potassium acetate (1.3 g, 13.2mmol), [1,1′-bis(diphenylphosphino)ferrocene)dichloropalladium(II) (0.24g, 0.3 mmol) and 37 mL of 1,4-dioxane are combined in a pressure vessel.The vessel is flushed with argon and sealed. The mixture is stirred at120° C. for 45 minutes and cooled to room temperature.

Step B: 2-Bromo-1,4-benzodioxine-5-carboxamide (0.9 g, 3.6 mmol),[1,1′-bis(diphenylphosphino)ferrocene)dichloropalladium(II) (0.12 g,0.17 mmol), and 2M aqueous sodium carbonate (3.3 mL, 6.6 mmol) are addedto the reaction mixture from step A. The vessel is flushed with argonand sealed. The mixture is stirred at 100° C. for 2 hours. The mixtureis filtered through diatomaceous earth and rinsed with 10% MeOH in DCM(150 mL). The filtrate is concentrated. The resulting crude product ispurified by silica gel flash column chromatography eluting with 0-6%MeOH in DCM as the gradient to afford 0.39 g of2-[5-[2-(dimethylamino)-2-oxo-ethyl]-3-pyridyl]-1,4-benzodioxine-5-carboxamide.

Step C: To a pre-degassed solution of2-[5-[2-(dimethylamino)-2-oxo-ethyl]-3-pyridyl]-1,4-benzodioxine-5-carboxamide(0.62 g, 1.8 mmol) in 49 mL of acetic acid is added 124 mg of 10 wt %palladium on carbon. The resulting mixture is evacuated and back-filledwith H₂ (repeated twice). The mixture is then hydrogenated for 2 hours.The mixture is filtered through diatomaceous earth and rinsed withEtOAc. The filtrate is concentrated. The resulting residue isre-dissolved in EtOAc. Saturated NaHCO₃ solution (20 mL) and water (10mL) are added. The two layers are separated. The aqueous layer isextracted with EtOAc (4×50 mL). The combined organic layers are driedover sodium sulfate, filtered, and concentrated. The resulting crudeproduct is purified by silica gel flash column chromatography elutingwith 0-10% MeOH in DCM to afford 0.43 g of2-[5-[2-(dimethylamino)-2-oxo-ethyl]-3-pyridyl]-2,3-dihydro-1,4-benzodioxine-5-carboxamide.The stereoisomers are separated by chiral SFC.

Compounds 30 through 32 in Table 1 are synthesized according to theprocedure for Example 3, substituting either commercially availablereagents or the appropriate intermediates described above.

Compound 33 in Table 1 is synthesized according to the procedure forExample 3, substituting 2-bromo-benzo[1,4]dioxine-5-carboxylic acidmethylamide for 2-bromo-1,4-benzodioxine-5-carboxamide in Step B.

Example 4:2-[5-(2-Oxo-oxazolidin-3-ylmethyl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carbonitrile(Cpd 34, Table 1)

To a solution of2-[5-(2-oxo-oxazolidin-3-ylmethyl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid amide, compound 31, enantiomer A (35 mg, 0.10 mmol) in 2.0 mL of1,4-dioxane is added pyridine (0.16 mL, 1.97 mmol) followed bytrifluoroacetic anhydride (0.14 mL, 0.98 mmol). After 5 minutes, thereaction is poured into 7.5 mL of water and 7.5 mL of saturated NaHCO₃solution. The product is extracted into EtOAc (2×) and the combinedorganics are washed once with water and then dried (MgSO₄). The organicis filtered and concentrated to give the crude product which is purifiedvia flash column chromatography on a Biotage KP-NH column eluting withmethanol in DCM to afford 25 mg of the title compound.

Compounds 35 through 43 in Table 1 are synthesized according to theprocedure for Example 4, substituting the appropriate compoundsdescribed above. Chiral SFC is utilized for enantiomer resolution forexamples synthesized from racemic starting materials, and conditions canbe found in Table 2. All other examples are prepared fromenantiomerically pure starting materials.

Compound 44 in table 1 is synthesized according to the procedures inExample 2 and Example 4, substituting the appropriate intermediatedescribed above.

Example 5:2-(5-Ethoxy-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid amide (Cpd 45, Table 1)

Step A: 2-(5-Benzyloxy-pyridin-3-yl)-benzo[1,4]dioxine-5-carboxylic acidmethyl ester is synthesized from 3-benzyloxy-5-bromo-pyridine and2-bromo-benzo[1,4]dioxine-5-carboxylic acid methyl ester according tothe method of Example 2, steps A and B.

Step B: 2-(5-Benzyloxy-pyridin-3-yl)-benzo[1,4]dioxine-5-carboxylic acidmethyl ester (500 mg, 1.3 mmol) is dissolved in 10 mL of DCM and 10 mLof methanol. Then 5% Pd on carbon (280 mg, 0.13 mmol) is added. Ahydrogen balloon is attached to the reaction flask and the mixture isstirred under hydrogen atmosphere for 1.5 hours. Then the mixture isfiltered and the filtrate is concentrated to give 375 mg of2-(5-hydroxy-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester.

Step C: Ethanol (0.041 mL, 0.70 mmol),2-(5-hydroxy-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester (100 mg, 0.35 mmol) and triphenylphosphine (180 mg,0.70 mmol) are dissolved in 3.0 mL of THF and diisopropylazodicarboxylate (0.14 mL, 0.70 mmol) is added. The mixture is stirredfor 5 hours and the solvent is removed. The residue is purified by flashcolumn chromatography on silica gel to give 79 mg of2-(5-ethoxy-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester.

Step D: Lithium hydroxide monohydrate (21 mg, 0.50 mmol) is dissolved in1.0 mL of water and this solution is added into2-(5-ethoxy-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester (79 mg, 0.25 mmol) solution in 2.0 mL of 1,4-dioxane.The mixture is stirred for 64 hours and 0.3 mL of acetic acid is added.Then all the solvents are removed and 25 mL of water is added. A solidis formed and it is filtered, rinsed with more water and dried to give71 mg of2-(5-ethoxy-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid.

Step E:2-(5-Ethoxy-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid (71 mg, 0.24 mmol) is dissolved in 2.0 mL of DMF and1,1′-carbonyldiimidazole (77 mg, 0.48 mmol) is added. The mixture isheated at 60° C. for 1 hour and it is then cooled down to roomtemperature. Then 28% ammonium hydroxide aqueous solution (0.33 mL, 2.4mmol) is added and the mixture is stirred for another hour. Then 25 mLof water is added and a solid is formed. The solid is filtered, rinsedwith more water and dried to give 53 mg of the titled product.Enantiomers of the titled compound are separated using chiral SFC.

Compounds 46 and 47 in Table 1 are synthesized according to theprocedure for Example 5, substituting ethanol in Step C with theappropriate commercially available alcohols.

Example 6:2-[5-(1-isobutyryl-piperidin-4-yloxy)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid amide (Cpd 48, Table 1)

Step A:4-[5-(5-Methoxycarbonyl-2,3-dihydro-benzo[1,4]dioxin-2-yl)-pyridin-3-yloxy]-piperidine-1-carboxylicacid tert-butyl ester is synthesized according to Example 5, Step A toStep C, substituting ethanol in Step C with commercially available4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester.

Step B:4-[5-(5-Methoxycarbonyl-2,3-dihydro-benzo[1,4]dioxin-2-yl)-pyridin-3-yloxy]-piperidine-1-carboxylicacid tert-butyl ester (380 mg, 0.80 mmol) is dissolved in 5.0 mL of DCMand 1.0 mL of trifluoroacetic acid is added. The mixture is stirred for2 hours and all the solvent is removed. EtOAc (30 mL) is added alongwith 10 mL of saturated aqueous solution of NaHCO₃. The mixture isstirred for 10 min and the aqueous layer is separated and extracted withEtOAc. The organic layers are combined and concentrated to give 300 mgof2-[5-(piperidin-4-yloxy)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester.

Step C:2-[5-(Piperidin-4-yloxy)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester (300 mg, 0.80 mmol) is dissolved in 5.0 mL of DCM.Then isobutyryl chloride (0.16 mL, 1.52 mmol) and triethyl amine (0.28mL, 2.04 mmol) are added. After the mixture is stirred for 16 hours, 5mL of saturated aqueous solution of NaHCO₃ (5 mL) is added along with 15mL of water and 15 mL of DCM. The mixture is stirred for 10 minutes andthe aqueous layer is separated and extracted with DCM. The organiclayers are combined and concentrated to give the crude product.Purification by flash column chromatography on silica gel affords 160 mgof2-[5-(1-isobutyryl-piperidin-4-yloxy)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester.

Step D: Lithium hydroxide monohydrate (30 mg, 0.73 mmol) is dissolved in1.0 mL of water and this solution is added into2-[5-(1-isobutyryl-piperidin-4-yloxy)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester (160 mg, 0.36 mmol) solution in 2.0 mL of 1,4-dioxane.The mixture is stirred for 64 hours and 3 mL of acetic acid is addedalong with 20 mL of EtOAc and 20 mL of water. The aqueous layer isseparated and extracted with EtOAc. All the organic layers are combinedand concentrated to give 110 mg of2-[5-(1-isobutyryl-piperidin-4-yloxy)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid.

Step E:2-[5-(1-Isobutyryl-piperidin-4-yloxy)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid (110 mg, 0.26 mmol) is dissolved in 2.0 mL of DMF and1,1′-carbonyldiimidazole (84 mg, 0.52 mmol) is added. The mixture isheated at 60° C. for 1 hour and it is then cooled down to roomtemperature. Then 28% ammonium hydroxide aqueous solution (0.36 mL, 2.6mmol) is added and the mixture is stirred for another hour. Then 25 mLof water is added and a solid is formed. The solid is filtered, rinsedwith more water and dried to give 75 mg of the titled product.Enantiomers of the titled compound are separated using chiral SFC.

Example 7:2-[5-(2,2,2-Trifluoro-1-hydroxy-ethyl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid amide (Cpd 49, Table 1)

Step A:2-[5-(2,2,2-Trifluoro-1-hydroxy-ethyl)-pyridin-3-yl]-benzo[1,4]dioxine-5-carboxylicacid amide is prepared from1-(5-bromo-pyridin-3-yl)-2,2,2-trifluoro-ethanol and2-bromo-benzo[1,4]dioxine-5-carboxylic acid methyl ester according, toExample 2, Step A through Step C. Enantiomers are separated using.Chiral SEC (LUX Celluse-2, 30% (1:1:1 MeOH:EtOH:i-PrOH+0.1% DEA):CO₂, 70mL/min, 120 bar, 35° C.).

Step B:2-[5-(2,2,2-Trifluoro-1-hydroxy-ethyl)-pyridin-3-yl]-benzo[1,4]dioxine-5-carboxylicacid amide, enantiomer A (125 mg, 0.36 mmol) is hydrogenated accordingto Example 2, Step D to give 90 mg of product. Chiral SFC of thismaterial delivers 14 mg of 49AA and 14 mg of 49AB.

Step C:2-[5-(2,2,2-Trifluoro-1-hydroxy-ethyl)-pyridin-3-yl]-benzo[1,4]dioxine-5-carboxylicacid amide, enantiomer B (120 mg, 0.34 mmol) is hydrogenated accordingto Example 2, Step D to give 90 mg of product. Chiral SFC of thismaterial delivers 13 mg of 49BA and 15 mg of 49BB.

Example 8:2-[5-(4-Hydroxy-tetrahydro-pyran-4-yl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carbonitrile,(Cpd 50, Table 1)

A mixture of2-[5-(4-hydroxy-tetrahydro-pyran-4-yl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid amide, enantiomer B (40 mg, 0.1 mmol) and Palladium(II) chloride(20 mg, 0.1 mmol) in 1 mL of 1:1 ACN:Water is heated in a sealed vial at50° C. for 16 hours. The mixture is allowed to cool and water is added.The resultant precipitate is filtered off and dried. The solid isdissolved in 10% water in DMSO and is purified by prep HPLC. Fractionsare concentrated to dryness to provide 8 mg of the title compound.

Example 9:2-(7-Hydroxy-6,7-dihydro-5H-[2]pyrindin-4-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid amide (Cpd 52, Table 1)

Step A:2-(7-Hydroxy-6,7-dihydro-5H-[2]pyrindin-4-yl)-benzo[1,4]dioxine-5-carboxylicacid methyl ester is synthesized from4-bromo-6,7-dihydro-5H-[2]pyrindin-7-ol and2-bromo-benzo[1,4]dioxine-5-carboxylic acid methyl ester according toExample 2, Steps A and B, Enantiomers are separated using SFC (LUXCellulose-1, 45% (MeOH)CO₂, 125 mL/min, 120 bar, 40° C.).

Step B:2-(7-Hydroxy-6,7-dihydro-5H-[2]pyrindin-4-yl)-benzo[1,4]dioxine-5-carboxylicacid methyl ester, enantiomer A is converted into the title compound,according to Example 2, Steps C and D. Chiral SFC gives 52AA and 52AB.

Step C:2-(7-Hydroxy-6,7-dihydro-5H-[2]pyrindin-4-yl)-benzo[1,4]dioxine-5-carboxylicacid methyl ester, enantiomer B is converted into the title compound,according to Example 2, Steps C and D. Chiral separation using SFC gives52BA and 52BB.

Example 10:N-[5-(5-Cyano-2,3-dihydro-benzo[1,4]dioxin-2-yl)-pyridin-3-ylmethyl]-2,2,2-trifluoro-acetamide(Cpd 53, Table 1) and ethanesulfonic acid[5-(5-cyano-2,3-dihydro-benzo[1,4]dioxin-2-yl)-pyridin-3-ylmethyl]-amide(54, Table 1)

Step A: 2-Bromo-benzo[1,4]dioxine-5-carboxylic acid methyl ester and(5-bromo-pyridin-3-ylmethyl)-carbamic acid tert-butyl ester areconverted to[5-(5-carbamoyl-2,3-dihydro-benzo[1,4]dioxin-2-yl)-pyridin-3-ylmethyl]-carbamicacid tert-butyl ester according to Example 2, Step A through Step D.

Step B:[5-(5-Carbamoyl-2,3-dihydro-benzo[1,4]dioxin-2-yl)-pyridin-3-ylmethyl]-carbamicacid tert-butyl ester is converted to[5-(5-cyano-2,3-dihydro-benzo[1,4]dioxin-2-yl)-pyridin-3-ylmethyl]-carbamicacid tert-butyl ester according to Example 4.

Step C:[5-(5-Cyano-2,3-dihydro-benzo[1,4]dioxin-2-yl)-pyridin-3-ylmethyl]-carbamicacid tert-butyl ester (140 mg, 0.4 mmol) is dissolved in 5 mL of DCM.Trifluoroacetic acid (0.5 mL) is added and the reaction mixture isstirred at room temperature for 2 hours. The solvent is removed to give100 mg of2-(5-aminomethyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carbonitrile.Crude2-(5-aminomethyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carbonitrile(100 mg, 0.4 mmol) containing residual trifluoroacetic acid is dissolvedin 5 mL of THF. N,N-Diisopropylethylamine (0.12 mL, 0.8 mmol) andethanesulfonyl chloride (75 μL, 0.8 mmol) are added. The reactionmixture is stirred at room temperature for two hours. It is thenconcentrated to dryness and is purified by flash chromatography on aBiotage KP-NH column eluting with EtAOc in heptanes to give 26 mg of 53and 40 mg of ethanesulfonic acid[5-(5-cyano-2,3-dihydro-benzo[1,4]dioxin-2-yl)-pyridin-3-ylmethyl]-amide(54). Enantiomers of 54 are separated using chiral SFC.

Example 11:2-{5-[2-((R)-3-Hydroxy-pyrrolidin-1-yl)-2-oxo-ethyl]-pyridin-3-yl}-2,3-dihydro-benzo[1,4]dioxine-5-carbonitrile(Cpd 55, Table 1)

Step A: Trifluoro-methanesulfonic acid(R)-3-{2-[5-(5-cyano-2,3-dihydro-benzo[1,4]dioxin-2-yl)-pyridin-3-yl]-acetyl}-cyclopentylester is prepared from2-(5-bromo-pyridin-3-yl)-1-((R)-3-hydroxy-pyrrolidin-1-yl)-ethanone and2-bromo-benzo[1,4]dioxine-5-carboxylic acid methyl ester according toExample 2, Step A through Step D and Example 4.

Step B: To Trifluoro-methanesulfonic acid(R)-3-{2-[5-(5-cyano-2,3-dihydro-benzo[1,4]dioxin-2-yl)-pyridin-3-yl]-acetyl}-cyclopentylester (140 mg, 0.30 mmol) in 5 mL of 1:1 THF:water is added lithiumhydroxide (72 mg, 3.0 mmol). The reaction is stirred at room temperaturefor 16 hours. The reaction is concentrated to dryness and the residue ispartitioned between EtOAc and water. The EtOAc layer is concentrated todryness and the residue is purified by silica gel chromatography to give90 mg of the racemic title compound. Enantiomers of 55 are separatedusing chiral SFC.

Example 12:2-[5-Fluoro-4-((S)-1-hydroxy-ethyl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid amide (Cpd 56, Table 1)

Step A: 3-Bromo-5-fluoro-pyridine (13 g, 74 mmol) is dissolved in 140 mLof dry THF and cooled down to −78° C. LDA solution (44 mL, 2.0 M in THF,88 mmol) is added and the mixture is stirred for 2 hours at −78° C. Thenacetaldehyde solution (30 mL, 5.0 M in THF, 150 mmol) is added at −78°C. and the reaction is continued for another 30 minutes. Then saturatedaqueous NH₄Cl solution (200 mL) is added and the mixture is warmed up toroom temperature. EtOAc (100 mL) is added along with 75 mL of water. Theaqueous layer is separated and extracted with EtOAc (2×75 mL). Theorganic layers are combined and concentrated to give the crude product.Purification by flash column chromatography affords 14 g of the racemicproduct. Chiral separation of the racemic product using chiral SFCaffords 6.5 g of (R)-1-(3-bromo-5-fluoro-pyridin-4-yl)-ethanol and 6.4 gof (S)-1-(3-bromo-5-fluoro-pyridin-4-yl)-ethanol.

Step B: A solution of (S)-1-(3-bromo-5-fluoro-pyridin-4-yl)-ethanol(1.62 g, 7.4 mmol) in 25 mL of THF is cooled to 0° C. and 60% sodiumhydride (736.23 mg, 18.4 mmol) is then added. The mixture is stirred at0° C. for 1 hour then cooled to −78° C. n-Butyllithium 1.08 M in hexanes(10.23 mL, 11.0 mmol) is added, followed by triisopropyl borate (2.55mL, 11.0 mmol). The cooling bath is removed and the mixture is stirredat room temperature for 16 hours. The reaction mixture is cooled to 0°C. and then quenched with 5.0 mL of 1:1 solution of conc. H₂SO₄:water.The mixture is stirred at room temperature for 1 hour. The organicsolvent is evaporated and the aqueous layer is then neutralized to pH6-7. The aqueous layer is extracted with EtOAc (3×) and the combinedorganic layers are washed with brine, dried over MgSO₄, filtered andconcentrated to give crude(S)-4-fluoro-3-methyl-2-oxa-6-aza-1-bora-indan-1-ol.

Step C: 2-Bromo-benzo[1,4]dioxine-5-carboxylic acid methyl ester (1.0 g,3.7 mmol), crude (5)-4-fluoro-3-methyl-2-oxa-6-aza-1-bora-indan-1-ol(924 mg, 5.5 mmol),[1,1′-bis(diphenylphosphino)ferrocene)dichloropalladium(II) DCM complex(150.63 mg, 0.18 mmol), 1,4-dioxane (15.00 ml) and 2.0M Na₂CO₃ aqueoussolution (3.69 mL, 7.4 mmol) are added to a pressure vessel. The vesselis flushed with argon, sealed and stirred at 100° C. for 2 hours. Thereaction mixture is cooled to room temperature, diluted with EtOAc and25 mL of water and the mixture is filtered on diatomaceous earth. Thelayers of the filtrate are separated and the organic layer is washedwith brine, dried over MgSO₄, filtered and concentrated. Purification bysilica gel flash column chromatography eluting with 50-100%EtOAc/heptane) to give 659 mg of2-[5-fluoro-4-((S)-1-hydroxy-ethyl)-pyridin-3-yl]-benzo[1,4]dioxine-5-carboxylicacid methyl ester.

Step D:2-[5-Fluoro-4-((S)-1-hydroxy-ethyl)-pyridin-3-yl]-benzo[1,4]dioxine-5-carboxylicacid methyl ester is converted to the title compound according toExample 2, Steps C and D. The benzodiozane enantiomers are separated bychiral SFC.

Compound 57 in Table 1 is synthesized according to the procedure forExample 12, substituting (R)-1-(3-bromo-5-fluoro-pyridin-4-yl)-ethanolin Step B.

Example 13:2-[5-Fluoro-4-(1-hydroxy-1-methyl-ethyl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid amide (Cpd 58, Table 1)

Step A: To a solution of2-[5-fluoro-4-(1-hydroxy-ethyl)-pyridin-3-yl]-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid amide (cpd 57) (632.00 mg, 2.0 mmol), and Dess-Martin periodinane(1.01 g, 2.4 mmol) in 35 mL of acetonitrile is added trifluoroaceticacid (0.15 mL, 2.0 mmol). The heterogenous mixture is stirred at roomtemperature for 2 days. The reaction mixture is diluted with 150 mL of15% MeOH/DCM. 1N NaOH and 2M Na₂S₂O₃ are added and the mixture isfiltered. The layers are separated, and the organic layer isconcentrated. Purification by silica gel flash column chromatographyeluting with 50-100% EtOAc/heptane gives 550 mg of2-(4-acetyl-5-fluoro-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid amide.

Step B: A solution of2-(4-acetyl-5-fluoro-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid amide (440.00 mg, 1.4 mmol) in 44 mL of THF is cooled to 0° C. andthen treated with 2.0 M methylmagnesium bromide solution in THF (2.3 mL,6.6 mmol). The mixture is stirred at 0° C. for 30 minutes. The reactionmixture is quenched with saturated aqueous NH₄Cl solution and dilutedwith EtOAc/water. The aqueous layer is separated, and back-extractedwith EtOAc. The combined organic layers are washed with brine, driedover MgSO₄, filtered and concentrated. Purification by columnchromatography on a Biotage KP-NH column eluting with 50-100% EtOAcgives 125 mg of the title compound. Enantiomers are separated usingchiral SFC.

Example 14:2-[5-(Cyclopropyl-ethanesulfonylamino-methyl)-pyridin-3-yl]-benzo[1,4]dioxine-5-carboxylicacid amide (Cpd 60, Table 1)

Step A: To a mixture of 5-bromonicotinaldehyde (0.50 g, 2.69 mmol) andethanesulfonamide (0.37 g, 3.36 mmol) in 9.0 mL of toluene is addedtitanium(IV) isopropoxide (1.59 mL, 5.4 mmol). The reaction mixture isstirred at 120° C. for 3 hours after which time it is concentrated todryness. The remaining residue is dissolved in 10 mL of THF and cooledto −40° C. Cyclopropylmagnesium bromide (16.13 mL, 8.1 mmol) is addeddrop-wise and the reaction mixture is allowed to gradually warm to roomtemperature. After 16 hours, the reaction mixture is diluted with EtOAcand washed with saturated aqueous NH₄Cl solution then brine. The organiclayer is dried (MgSO₄), filtered and concentrated. The remaining residueis purified via silica gel flash column chromatography eluting with 0-5%MeOH/DCM to give 0.59 g of ethanesulfonic acid[(5-bromo-pyridin-3-yl)-cyclopropyl-methyl]amide.

Step B: Ethanesulfonic acid[(5-bromo-pyridin-3-yl)-cyclopropyl-methyl]-amide and2-bromo-benzo[1,4]dioxine-5-carboxylic acid methyl ester are convertedto2-[5-(cyclopropyl-ethanesulfonylamino-methyl)-pyridin-3-yl]-benzo[1,4]dioxine-5-carboxylicacid amide according to Example 2, Steps A-C. Enantiomers are separatedusing SEC (Regis S,S) Whelk-O 1, 40% (EtOH+1% Isopropylamine):CO₂, 80mL/min, 100 bar, 25° C.).

Step C:2-[5-(Cyclopropyl-ethanesulfonylamino-methyl)-pyridin-3-yl]-benzo[1,4]dioxine-5-carboxylicacid amide, enantiomer A is hydrogenated according to Example 2, Step D.Chiral SFC yields 60AA and 60AB.

Step D:2-[5-(Cyclopropyl-ethanesulfonylamino-methyl)-pyridin-3-yl]-benzo[1,4]dioxine-5-carboxylicacid amide, enantiomer B is hydrogenated according to Example 2, Step D.Chiral SFC yield 60BA and 60BB.

Example 15:2-(5-Cyano-4-methyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid amide (Cpd 61, Table 1)

Step A: To a stirred suspension of 5-bromo-4-methyl-nicotinic acid (1.75g, 8.10 mmol) in 20 mL of DMF is added CDI (1.97 g, 12.2 mmol). Themixture is warmed at 65° C. for 0.75 hour after which time it is cooledto room temperature and treated with ammonium hydroxide (10.1 ml, 81.0mmol). After stirring for 2 hours the reaction is poured into water (150ml) and the product is extracted into EtOAc (3×). The combined organicsare dried (MgSO₄), filtered and concentrated. The crude residue ispurified via silica gel flash column chromatography eluting with 0-6%MeOH/DCM to afford 1.4 g of 5-bromo-4-methyl-nicotinamide.

Step B: 5-Bromo-4-methyl-nicotinamide and2-bromo-benzo[1,4]dioxine-5-carboxylic acid methyl ester are convertedto2-(5-carbamoyl-4-methyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester according to Example 1, Steps A-C

Step C: To a stirred solution of2-(5-carbamoyl-4-methyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester (180 mg, 0.55 mmol) in 10.0 mL of 1,4-dioxane andpyridine (0.89 ml, 10.9 mmol) is added trifluoroacetic anhydride (0.77ml, 5.5 mmol) in a drop-wise manner over 10 minutes. Upon completeaddition the reaction is stirred for 5 minutes after which time it ispoured into water and NaHCO₃ (sat., 1:1, 150 mL). The mixture is dilutedwith EtOAc and the layers are separated. The organic layer is washedonce with water and then dried (MgSO₄). Filtration and concentrationgave 160 mg of2-(5-cyano-4-methyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester.

Step D: A suspension of2-(5-cyano-4-methyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester (160 mg, 0.52 mmol) in 7N ammonia in methanol (5.0 ml,35.0 mmol) is warmed to 85° C. After 24 hours the reaction is cooled toroom temperature and concentrated. The remaining crude is purified viaflash column chromatography on a Biotage KP-NH column eluting with DCMto give 70 mg of the title compound. Enantiomers were separated usingSFC.

Example 16:2-(5-{[Imino(methyl)oxo-λ⁶-sulfanyl]methyl}pyridine-3-yl)-2,3-dihydro-1,4-benzodioxine-5-carbonitrile(Cpd 62, Table 1)

Step A:(2-(5-Hydroxymethyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester is synthesized from (5-bromo-pyridin-3-yl)-methanoland 2-bromo-benzo[1,4]dioxine-5-carboxylic acid methyl ester accordingto Example 1, steps A through C. Enantiomers are separated by chiral SFC(Chiracel-OJ-H, 0.5% DEA in methanol, 100 mL/min, 100 bar, 25° C.).

Step B: To a cooled (0° C.) solution of(2-(5-hydroxymethyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester, enantiomer B (1.80 g, 6.0 mmol) andtriphenylphosphine (1.88 g, 7.2 mmol) in 50 mL of DCM is added carbontetrabromide (2.38 g, 7.2 mmol). The reaction is stirred for 30 minutesafter which time the mixture is concentrated in vacuo. The crude residueis purified by silica gel flash column chromatography eluting with10-100% EtOAc/heptane to give 1.3 g of2-(5-bromomethyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester, enantiomer B.

Step C: A solution of2-(5-bromomethyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester, enantiomer B (1.3 g, 3.6 mmol) in 35 mL of DMF istreated with sodium thiomethoxide (325 mg, 4.6 mmol) and potassiumcarbonate (987 mg, 7.1 mmol) and stirred at room temperature overnight.After this time the reaction is filtered and the solids are washed withDCM. The combined filtrates are concentrated and the remaining residueis purified via silica gel flash column chromatography eluting with 0-8%MeOH in DCM to give 1 g of2-(5-methylsulfanylmethyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester, enantiomer B.

Step D: To a 0° C. stirred solution of2-(5-methylsulfanylmethyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester, enantiomer B (1.00 g, 3.0 mmol) in 45 mL ofchloroform is added 3-chloroperoxybenzoic acid (593 mg, 2.4 mmol) in 4additions over 20 minutes. The reaction is stirred for 15 minutes at 0°C. and treated with triethylamine (1.5 ml) and concentrated. Theremaining residue is purified via flash chromatography on a BiotageKP-NH column eluting with 10-100% EtOAc/heptane to give 600 mg of2-(5-methanesulfinylmethyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester, mixture of diastereomers BA and BB.

Step E: To a solution of2-(5-methanesulfinylmethyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester, diastereomers BA and BB (600 mg, 1.7 mmol) in 30 mLof DCM is sequentially added 2,2,2-trifluro-acetamide (390 mg, 3.5mmol), magnesium oxide (278 mg, 6.9 mmol), rhodium(II) acetate dimer (53mg, 0.1 mmol), and iodobenzene diacetate (835 mg, 2.6 mmol). Thereaction is allowed to stir at room temperature for 17 hours. After thistime, the reaction is re-charged with the reactants using the originalequivalents. After stirring overnight at room temperature the reactionis filtered and the solids are washed with DCM. The combined filtratesare concentrated and the remaining crude residue is purified via silicagel flash column chromatography eluting with 5-100% EtOAc/heptane togive 160 mg of methyl2-(5-{[methyl(oxo)[(trifluoroacetyl)imino]-λ⁶-sulfanyl]methyl}pyridine-3-yl)-2,3-dihydro-1,4-benzodioxine-5-carboxylate,mixture of diastereomers BA and BB.

Step F: A 20 mL microwave reaction vessel is charged with methyl2-(5-{[methyl(oxo)[(trifluoroacetyl)imino]-λ⁶-sulfanyl]methyl}pyridine-3-yl)-2,3-dihydro-1,4-benzodioxine-5-carboxylate,diasteromers BA and BB (160 mg, 0.4 mmol) and 7N ammonia in methanol (8mL). The vessel is capped and warmed at 85° C. for 2 days. After thistime the reaction is cooled and concentrated. The crude is purified viaHPLC (5-60% ACN/H2O, 20 minutes, TFA modified solvents) and theproduct-containing fractions are concentrated to give2-(5-{[imino(methyl)oxo-λ⁶-sulfanyl]methyl}pyridine-3-yl)-2,3-dihydro-1,4-benzodioxine-5-carboxamide,mixture of diastereomers BA and BB.

Step G: To a mixture of2-(5-{[imino(methyl)oxo-λ⁶-sulfanyl]methyl}pyridine-3-yl)-2,3-dihydro-1,4-benzodioxine-5-carboxamide,diasteromers BA and BB (220 mg, 0.5 mmol) in 25 mL of dioxane is addedpyridine (0.77 mL, 9.5 mmol) and trifluoroacetic anhydride (0.67 mL, 4.8mmol) drop-wise. The reaction is stirred at room temperature for 15minutes after which time the reaction is concentrated to dryness. Theremaining residue is treated with 7N MeOH in ammonia (50 mL) and themixture is concentrated. The remaining residue is purified via HPLC(10-100% CH₃CN/H₂O over 20 minutes, 0.1% TFA) to give 70 mg of2-(5-{[imino(methyl)oxo-λ⁶-sulfanyl]methyl}pyridine-3-yl)-2,3-dihydro-1,4-benzodioxine-5-carbonitrile.Diastereomers BA and BB are resolved by chiral SFC to yield 62BA and62BB.

Step H:(2-(5-Hydroxymethyl-pyridin-3-yl)-2,3-dihydro-benzo[1,4]dioxine-5-carboxylicacid methyl ester, enantiomer A is converted into2-(5-{[imino(methyl)oxo-λ⁶-sulfanyl]methyl}pyridine-3-yl)-2,3-dihydro-1,4-benzodioxine-5-carbonitrile,mixture of diasteromers AA and AB according to the above procedure,Example 16, Steps B through G. diasteromers AA and AB are resolved bychiral SFC to yield 62AA and 62BB.

TABLE 2 Chiral SFC Separation Conditions Flow Rate Pressure Temp Cpd #Column Mobile Phase (mL/min) (bar) (° C.)  1 ChiralPak IC 32%(1:1:1MeOH:EtOH:IPA + 0.1% 85 110 40 DEA):CO₂  2 RegisPack 30% (2:1:1MeOH:EtOH:IPA):CO₂ 130 120 35  3 RegisPack 45%(EtOH):CO₂ 125 120 35  4ChiralPak IC 31%(1:1:1 MeOH:EtOH:IPA + 1% DEA):CO₂ 84 130 40  5RegisPack 25%(1:1:1MeOH:EtOH:IPA):CO₂ 75 125 40  6A LUX Cellulose-325%(1:1:1MeOH:EtOH:IPA):CO₂ 155 120 35  6B LUX Cellulose-3 25%(1:1:1MeOH:EtOH:IPA):CO₂ 150 120 35  7 LUX Cellulose-1 27%(1:1:1MeOH:EtOH:IPA):CO₂ 85 120 40  8 ChiralPak IC 40% (MeOH):CO₂ 85 120 40  9LUX Cellulose-1 28%(1:1:1 MeOH:EtOH:IPA):CO₂ 145 120 40 10 ChiralPakAD-H 32% (MeOH):CO₂ 90 120 40 11 RegisPack 25% (1:1:1 MeOH:EtOH:IPA):CO₂110 130 40 12 LUX Cellulose-3 18%(1:1:1MeOH:EtOH:IPA + 0.1% DEA):CO₂ 70120 35 13 LUX Cellulose-1 32%(1:1:1 MeOH:EtOH:IPA + 0.1% DEA):CO₂ 85 12035 14 ChiralPak IA 33%(1:1:1 MeOH:EtOH:IPA + 0.1% DEA):CO₂ 110 120 35 15LUX Cellulose-1 30%((1:1:1MeOH:EtOH:IPA) + 0.1% DEA): 90 120 35 CO₂ 16LUX Cellulose-3 40%(1:1:1 MeOH:EtOH:IPA):CO₂ 135 120 35 17 LUXCellulose-3 20%(1:1:1 MeOH:EtOH:IPA):CO₂ 140 120 35 18 LUX Cellulose-135% (1:1:1 MeOH:EtoH:IPA):CO₂ 140 120 35 19 LUX Cellulose-1 35%3:1:1(MeOH:EtOH:IPA):CO₂ 80 120 35 21 LUX Cellulose-1 25% (1:1:1MeOH:EtOH:IPA):CO₂ 90 130 40 23 LUX Cellulose-1 28%(6:7:7MeOH:EtOH:IPA):CO₂ 80 120 40 24 ChiralPak AD-H 65% (1:1 MeOH:IPA +0.2% 65 100 25 isopropylamine):CO₂ 25 LUX Cellulose-1 30% (67%MeOH:33%(1:1EtOH:IPA) + 0.1% DEA): 65 120 35 CO₂ 26 RegisPack 25%(MeOH):CO₂ 70 120 40 27 LUX Cellulose-2 40% (MeOH):CO₂ 80 120 35 28ChiralPak IC 16%(1:1:1 MeOH:EtOH:IPA):CO₂ 90 120 40 29 RegisPack15%(11:5:4 MeOH:EtOH:IPA):CO₂ 90 120 40 30 RegisPack 25%(2:1IPA:MeOH)CO₂ 85 120 40 32 LUX Cellulose-1 30% (4:3:3 MeOH:EtOH:IPA):CO₂80 120 40 33 LUX Cellulose-3 15%(1:1:1MeOH:EtOH:IPA):CO₂ 115 120 40 36LUX Cellulose-3 25%(1:1:1 MeOH:EtOH:IPA):CO₂ 120 120 40 43 LUXCellulose-3 28%(1:1:1 MeOH:EtOH:IPA):CO₂ 115 120 40 44 LUX Cellulose-320%(85:15 MeOH:IPA):CO₂ 85 130 40 45 LUX Cellulose-130%(1:1:1MeOH:EtOH:IPA):CO₂ 110 140 40 48 LUX Cellulose-3 20%1:1:1(MeOH:EtOH:IPA):CO₂ 80 120 40 49A ChiralPak IC 25%1:1:1(MeOH:EtOH:IPA):CO₂ 120 120 35 49B ChiralPak IC 25%1:1:1(MeOH:EtOH:IPA):CO₂ 120 120 35 51 LUX Cellulose-1 40%(1:1MeOH:IPA):CO₂ 120 120 40 52A RegisPack 25% (6:3:1 IPA:MeOH:EtOH):CO₂ 80120 40 52B RegisPack 25% (1:1:1 MeOH:EtOH:IPA):CO₂ 80 120 40 54 LUXCellulose-3 20%(2:3:3 MeOH:EtOH:IPA):CO₂ 65 130 40 55 LUX Cellulose-325%(2:1:1 MeOH:EtOH:IPA):CO₂ 85 120 40 56 Chiracel OD-H 30% (1:1:1MeOH:EtOH:IPA):CO₂ 115 120 35 57 ChiralPak IA 30% (1:1:1MeOH:EtOH:IPA):CO₂ 120 120 35 58 LUX Cellulose-1 22% (1:1:1MeOH:EtOH:IPA):CO₂ 80 120 40 59 ChiralPak AD-H 45% (3:1 ACN:MeOH + 0.2%80 100 25 Isopropylamine):CO₂ 60A RegisPack 35% 1:1:1(MeOH:EtOH:IPA):CO₂ 145 120 35 60B RegisPack 30% (1:1:1MeOH:EtOH:IPA):CO₂ 140 120 35 61 ChiralPak AD-H 26%(1:1:1MeOH:EtOH:IPA):CO₂ 90 120 40 62A RegisPack 45%(MeOH):CO₂ 80 120 40 62BLUX Cellulose-4 31% (65:35 MeOH:IPA):CO₂ 70 140 40

TABLE 3 LC/MS Methods Mobile Mobile Flow Method Phase A Phase B Gradient(mL/min.) Column Time (min) % A % B A 0.1% Formic 0.1% 0 90.0 10.0 0.5Thermo Acid in Formic Acid in 0.5 90.0 10.0 Scientific, WaterAcetonitrile 1.5 1.0 99.0 Aquasil C18, 2.5 1.0 99.0 50 × 2.1 mm, 3.390.0 10.0 5μ 4.0 90.0 10.0 B 95% Water Acetonitrile + 90% A to 100% B in1.19 minutes, 0.8 BEH 5% Acetonitrile + 0.05% hold at 100% B to 1.70minutes 2.1 × 50 mm 0.05% Formic Acid C18, 1.7 μm Formic Acid particlediameter C 95% Water Acetonitrile 90% A to 100% B in 1.19 minutes 0.8BEH 5% Acetonitrile + hold at 100% B to 1.70 minutes 2.1 × 50 mm 2.5 mMC18, 1.7 μm Ammonium particle Bicarbonate diameter D 95% WaterAcetonitrile 90% A to 100% B in 4.45 minutes 0.8 BEH 5% Acetonitrile +hold at 100% B to 4.58 minutes 2.1 × 50 mm 2.5 mM C18, 1.7 μm Ammoniumparticle Bicarbonate diameter E 95% Water Acetonitrile + 95% A to 100% Bin 3.65 minutes, 0.6 HSS T3 5% Acetonitrile + 0.05% hold at 100% B to4.95 minutes 2.1 × 100 mm, 0.05% Formic Acid 1.8 μm particle Formic Aciddiameter F 95% Water Acetonitrile + 100% A hold for 1.00 minute, 0.6 HSST3 5% Acetonitrile + 0.05% 100% A to 95% B in 4.50 minutes 2.1 × 100 mm,0.05% Formic Acid hold at 100% B to 4.91 minutes 1.8 μm particle FormicAcid diameter

TABLE 4 LC/MS Data Mass Retention LCMS Cpd No Found Time (Min) Method 1A 357.2 0.37 B  1B 357.2 0.37 B  2A 322.4 0.61 B  2B 321.9 0.62 B  3A404.0 0.57 C  3B 404.0 0.56 C  4A 257.1 1.66 A  4B 257.2 1.66 A  5A354.2 1.23 A  5B 354.2 1.23 A  6AA 354.2 0.44 B  6AB 353.9 0.44 B  6BA354.0 0.44 B  6BB 354.0 0.44 B  7A 398.0 0.53 B  7B 397.9 0.53 B  8A349.2 1.16 A  8B 349.2 1.16 A  9A 325.0 0.76 B  9B 325.0 0.76 B 10A341.4 0.48 B 10B 340.9 0.47 B 11A 355.1 0.56 B 11B 355.1 0.56 B 12A398.2 0.53 B 12B 397.8 0.51 B 13A 384.3 1.14 A 13B 384.3 1.14 A 14A354.0 0.5 B 14B 354.0 0.5 B 15A 369.6 0.42 B 15B 369.7 0.42 B 16A 412.11.19 A 16B 412.1 1.19 A 17A 383.8 0.51 C 17B 384.0 0.52 C 18A 384.0 0.52C 18B 384.0 0.52 C 19A 370.2 0.5 C 19B 369.9 0.5 C 20A 286.9 0.43 C 20B286.9 0.43 C 21A 289.3 0.66 B 21B 289.0 0.66 B 22A 307.3 0.65 B 22B307.3 0.63 C 23A 270.9 1.15 A 23B 271.1 1.16 A 24A 368.0 0.57 C 24B368.0 0.57 C 25A 372.2 0.49 C 25B 372.2 0.49 C 26A 327.2 0.48 B 26B327.2 0.47 B 27A 315.7 2.04 F 27B 315.5 2.12 F 28A 339.2 1.32 A 28B339.2 1.32 A 29A 342.0 0.48 C 29B 342.0 0.48 C 30A 418.0 0.61 C 30B417.9 0.61 C 31A 355.9 0.56 D 31B 355.9 0.59 D 32A 359.3 1.26 A 32B359.3 1.26 A 33A 412.0 0.58 C 33B 412.0 0.57 C 34A 338.0 0.64 C 34B338.0 0.61 C 35A 336.1 0.63 B 35B 336.0 0.66 B 36A 352.2 1.23 A 36B352.2 1.22 A 37A 331.0 0.64 B 37B 331.2 0.64 B 38A 380.3 1.31 A 38B380.3 1.31 A 39A 336.0 0.68 C 39B 336.0 0.68 C 40A 394.9 0.81 B 40B394.9 0.85 B 41A 386.2 0.74 B 41B 386.3 0.74 B 42A 354.0 1.21 D 42B354.0 1.19 D 43A 366.2 1.24 A 43B 366.2 1.24 A 44A 346.8 0.69 B 44B346.9 0.69 B 45A 300.8 0.6 B 45B 300.9 0.6 B 46A 343.2 0.54 B 46B 342.90.54 B 47A 356.9 0.58 B 47B 356.8 0.58 B 48A 426.3 0.66 B 48B 426.0 0.66B 49AA 354.9 0.59 B 49AB 354.9 0.59 B 49BA 354.9 0.6 B 49BA 354.9 0.6 B50B 339.2 0.59 B 51A 275.2 0.62 C 51B 275.2 0.62 C 52AA 313.1 0.48 C52AB 313.1 0.46 C 52BA 313.1 0.46 C 52BB 313.1 0.47 C 53 363.8 0.78 B54A 359.9 0.65 B 54B 359.9 0.65 B 55A 366.3 1.29 A 55B 366.3 1.28 A 56A319.2 1.2 A 56B 319.2 1.2 A 57A 319.2 1.2 A 57B 319.2 1.2 A 58A 332.90.7 C 58B 332.9 0.7 C 59A 270.9 0.62 C 59B 271.3 0.41 B 60AA 418.0 1.64E 60AB 418.0 1.62 E 60BA 418.0 1.65 E 60BB 418.0 1.63 E 61A 296.0 0.6 C61B 296.0 0.6 C 62AA 330.1 2.66 F 62AB 330.1 2.66 F 62BA 330.1 0.55 B62BB 330.1 0.55 B

ASSESSMENT OF BIOLOGICAL ACTIVITY

Preparation of Cynomolgus Adrenal Mitochondria

The aldosterone synthase and cortisol synthase inhibition assays employcynomolgus adrenal gland mitochondria as the source of aldosteronesynthase (CYP11B2) and cortisol synthase (CYP11B1). Mitochondria areprepared from frozen cynomolgus monkey adrenal glands according toMethod A described in by J. D. McGarry et al. (Biochem. J., 1983, 214,21-28), with a final resuspension in the AT buffer described in R.Yamaguchi et al. (Cell Death and Differentiation, 2007, 14, 616-624),frozen as aliquots in liquid nitrogen and stored at −80° C. until use.Activity of CYP11B2 and CYP11B1 in these preparations is defined as theamount of enzyme that generates 1 pmol of product in one hour under theconditions described.

Inhibition of Aldosterone Synthase

The compounds of the invention may be evaluated for aldosterone synthaseinhibition by the following assay:

Assays are performed in 96-well format in a final volume of 60 μL/well,containing 100 mM potassium phosphate, pH 7.4, 1% (v/v) DMSO, andadditionally, 2 μM of corticosterone and 50 units of CYP11B2 activity.Reactions are started by the addition of NADPH to 1 mM and allowed toproceed for 90 minutes at 37° C. Reactions are terminated by theaddition of 60 μL of MeCN containing an internal standard for massspectrometry. One hundred microliters are then transferred to a glassfilter plate and centrifuged at 570×g for 5 minutes and the filtrate iscollected. Reaction product aldosterone is quantified by massspectrometry. To determine the assay blank value (0% activity), NADPH isomitted from some reactions.

Dose dependent inhibition is quantified by the inclusion of compound atvarious concentrations. Maximum activity (100%) is defined by reactionscontaining NADPH, but without compound. Activities at each concentrationare expressed as a percentage of the maximum activity (y-axis) andplotted against concentration of compound (x-axis) and the concentrationcorresponding to 50% activity (IC₅₀) determined using the XLFitcurve-fitting program using a 4-parameter logistic model.

Inhibition of Cortisol Synthesis

Assays are performed as for aldosterone synthase except for the use of150 units of CYP11B1, 11-deoxycortisol as substrate and cortisolmeasured as product.

Representative compounds of the present invention were tested foractivity in the above assays. Preferred compounds have an IC₅₀<1,000 nMand more preferred compounds have an IC₅₀<100 nM in this assay. Asexamples, data for representative compounds from Table 1 are shown inTable 5. Data for individual enantiomers are indicated by separateentries for enantiomers A and B.

TABLE 5 Biological Data Cyp11B2 Cyp11B1 Inhibition Inhibition Cpd NoIC₅₀ (nM) IC₅₀ (nM)  1A 570 >100000  1B 33 24000  2A 120 60  2B 17 230 3A 140 >30000  3B 33 5600  4A 68 3100  4B 22 2500  5A 310 >30000  5B 3922000  6AA 26 2200  6AB 70 10000  6BA 190 >30000  6BB 1000 >30000 7A >30,000 >30000  7B 47 19000  8A >30,000 >30000  8B 28 13000  9A 1105200  9B 10 2200 10A 66 3400 10B 14 230 11A 180 24000 11B 19 4400 12A100 18000 12B 10 2000 13A >1000 >100000 13B 95 66000 14A 29 12000 14B660 >30000 15A 48 15000 15B 540 >30000 16A 24 5200 16B 17 54 17A840 >30000 17B 67 18000 18A 610 16000 18B 41 1100 19A 1400 >30000 19B 7024000 20A 110 8200 20B 17 6800 21A 5 2600 21B 11 5300 22A 62 2000 22B 112300 23A 7.5 1400 23B 13 2000 24A 18 7000 24B 61 25000 25A 830 >10000025B 60 15000 26A 48 1800 26B 14 670 27A 17 2800 27B 31 5000 28A2200 >30000 28B 180 6100 29A 44 22000 29B 520 >30000 30A 75 22000 30B550 24000 31A 52 >30000 31B 250 >30000 32A 73 6000 32B 20 350 33A7400 >30000 33B 280 >30000 34A 9 8500 34B 100 >30000 35A 8 530 35B 336200 36A 100 >30000 36B 14 5800 37A 61 25000 37B 12 8000 38A 66 1500038B 6 700 39A 11 7300 39B 220 >30000 40A 300 >30000 40B 70 11000 41A 11024000 41B 8 3900 42A 110 18000 42B 10 1300 43A 25 22000 43B 460 >3000044A 9 2200 44B 81 13000 45A 18 210 45B 5 280 46A 260 7000 46B 17 420047A 110 8500 47B 10 1200 48A 51 3100 48B 20 240 49AA 90 3200 49AB 171100 49BA 66 2500 49BA 15 1100 50B 8 3300 51A 140 1200 51B 38 20000 52AA37 4100 52AB 37 820 52BA 71 16000 52BB 42 13000 53 35 24000 54A — 2300054B — 14000 55A 26 11000 55B 240 >30000 56A 78 14000 56B 24 14000 57A 5016000 57B 59 15000 58A 190 23000 58B 180 3000 59A 24 1100 59B 4 890 60AA530 >30000 60AB 60 >30000 60BA 3200 >30000 60BB 81 >30000 61A 52 2500061B 44 22000 62AA 320 >30000 62AB 500 >30000 62BA 25 14000 62BB 53 18000

Methods of Therapeutic Use

In accordance with the invention, there are provided novel methods ofusing the compounds of formula (I). The compounds disclosed hereineffectively inhibit aldosterone synthase. The inhibition of aldosteronesynthase is an attractive means for preventing and treating a variety ofdiseases or conditions that can be alleviated by lowering levels ofaldosterone. Thus, the compounds are useful for the treatment ofdiseases and conditions as described in the Background section,including the following conditions and diseases:

Diabetic kidney disease including diabetic nephropathy;

Non-diabetic kidney disease including glomerulosclerosis,glomerulonephritis, IGA nephropathy, nephritic syndrome and focalsegmental glomerulosclerosis (FSGS);

Cardiovascular diseases including hypertension, pulmonary arterialhypertension, Conn's syndrome, systolic heart failure, diastolic heartfailure, left ventricular dysfunction, left ventricular stiffness andfibrosis, left ventricular filing abnormalities, arterial stiffness,atherosclerosis and cardiovascular morbidity associated with primary orsecondary hyperaldosteronism;

Adrenal hyperplasia and primary and secondary hyperaldosteronism.

These disorders have been well characterized in man, but also exist witha similar etiology in other mammals, and can be treated bypharmaceutical compositions of the present invention.

Accordingly, a compound of formula I according to any of the embodimentsdescribed herein or a pharmaceutically acceptable salt thereof may beused for the preparation of a medicament for treating a disease ordisorder mediated by aldosterone synthase, including diabeticnephropathy, glomerulosclerosis, glomerulonephritis, IGA nephropathy,nephritic syndrome focal segmental glomerulosclerosis (FSGS),hypertension, pulmonary arterial hypertension, Conn's syndrome, systolicheart failure, diastolic heart failure, left ventricular dysfunction,left ventricular stiffness and fibrosis, left ventricular filingabnormalities, arterial stiffness, atherosclerosis and cardiovascularmorbidity associated with primary or secondary hyperaldosteronism,adrenal hyperplasia and primary and secondary hyperaldosteronism.

For therapeutic use, the compounds of the invention may be administeredvia a pharmaceutical composition in any conventional pharmaceuticaldosage form in any conventional manner. Conventional dosage formstypically include a pharmaceutically acceptable carrier suitable to theparticular dosage form selected. Routes of administration include, butare not limited to, intravenously, intramuscularly, subcutaneously,intrasynovially, by infusion, sublingually, transdermally, orally,topically or by inhalation. The preferred modes of administration areoral and intravenous.

The compounds of this invention may be administered alone or incombination with adjuvants that enhance stability of the inhibitors,facilitate administration of pharmaceutical compositions containing themin certain embodiments, provide increased dissolution or dispersion,increase inhibitory activity, provide adjunct therapy, and the like,including other active ingredients. In one embodiment, for example,multiple compounds of the present invention can be administered.Advantageously, such combination therapies utilize lower dosages of theconventional therapeutics, thus avoiding possible toxicity and adverseside effects incurred when those agents are used as monotherapies.Compounds of the invention may be physically combined with theconventional therapeutics or other adjuvants into a singlepharmaceutical composition. Advantageously, the compounds may then beadministered together in a single dosage form. In some embodiments, thepharmaceutical compositions comprising such combinations of compoundscontain at least about 5%, but more preferably at least about 20%, of acompound of formula (I) (w/w) or a combination thereof. The optimumpercentage (w/w) of a compound of the invention may vary and is withinthe purview of those skilled in the art. Alternatively, the compounds ofthe present invention and the conventional therapeutics or otheradjuvants may be administered separately (either serially or inparallel). Separate dosing allows for greater flexibility in the dosingregime.

As mentioned above, dosage forms of the compounds of this invention mayinclude pharmaceutically acceptable carriers and adjuvants known tothose of ordinary skill in the art and suitable to the dosage form.These carriers and adjuvants include, for example, ion exchangers,alumina, aluminum stearate, lecithin, serum proteins, buffer substances,water, salts or electrolytes and cellulose-based substances. Preferreddosage forms include tablet, capsule, caplet, liquid, solution,suspension, emulsion, lozenges, syrup, reconstitutable powder, granule,suppository and transdermal patch. Methods for preparing such dosageforms are known (see, for example, H. C. Ansel and N. G. Popovish,Pharmaceutical Dosage Forms and Drug Delivery Systems, 5th ed., Lea andFebiger (1990)). Dosage levels and requirements for the compounds of thepresent invention may be selected by those of ordinary skill in the artfrom available methods and techniques suitable for a particular patient.In some embodiments, dosage levels range from about 1-1000 mg/dose for a70 kg patient. Although one dose per day may be sufficient, up to 5doses per day may be given. For oral doses, up to 2000 mg/day may berequired. As the skilled artisan will appreciate, lower or higher dosesmay be required depending on particular factors. For instance, specificdosage and treatment regimens will depend on factors such as thepatient's general health profile, the severity and course of thepatient's disorder or disposition thereto, and the judgment of thetreating physician.

What is claimed is:
 1. A compound of the formula I

wherein: R¹ is selected from —C(O)NH₂, —C(O)NH(CH₃) and —CN; R² is—(X)—R⁴, wherein —(X)— is a bond, —CH₂—, or —O—; and R⁴ is selected from—H; C₁₋₃alkyl, optionally substituted with one to four groups selectedfrom —F, —SO₂C₁₋₃alkyl, and —OH; halogen; —CN; —SO₂C₁₋₃alkyl;—C(O)N(C₁₋₃alkyl)₂, provided —(X)— is not —O—; —NHC(O)R⁵ or—N(CH₃)C(O)R⁵, provided that —(X)— is —CH₂— and wherein R⁵ is selectedfrom C₃₋₆cycloalkyl and C₁₋₃alkyl optionally substituted with one tothree —F groups; —NHSO₂C₁₋₃alkyl; —CH(cyclopropyl)NHSO₂C₁₋₃alkyl;—OCH₂C(O)N(C₁₋₃alkyl)₂, provided that —(X)— is —CH₂—; —S(═O)(═NH)CH₃,provided that —(X)— is —CH₂—; heterocyclyl selected fromtetrahydropyranyl, tetrahydrofuranyl, pyrrolidinyl,1,1-dioxo[1,2]-thiazine, morpholinyl, oxazolidinyl, piperidinyl,azetidinyl, wherein said heterocyclyl is optionally substituted with oneto three groups selected from —C(O)C₁₋₃alkyl, halogen, —OH, oxo andC₁₋₃alkyl; —C(O)-heterocyclyl, provided that —(X)— is —CH₂, wherein saidheterocyclyl is selected from morpholin-4-yl, pyrrolidin-1-yl andpiperidin-1-yl, optionally substituted with one or two groups selectedfrom —F and —OH; C₃₋₆cycloalkyl optionally substituted with —CN or —OH;and phenyl, optionally substituted with —SO₂NH₂; and R³ is H, orC₁₋₃alkyl optionally substituted with —OH; or R₂ and R³ together form anannelated five-membered cycloalkyl ring optionally substituted with —OH;or a salt or a stereoisomer thereof.
 2. The compound according to claim1, wherein: R¹ is —C(O)NH₂ or —CN; R² is —(X)—R⁴, wherein —(X)— is abond, and R⁴ is selected from —CH₃, —CF₃; —CHF₂; —CH₂OH; —CH(OH)CH₃;—CH(OH)CF₃; —F; —CN; heterocyclyl selected from tetrahydropyranyl andpyrrolidinyl, wherein said heterocyclyl is optionally substituted withone to three groups selected from C₁₋₃alkyl, halogen, —OH and oxo;C₃₋₆cycloalkyl optionally substituted with —CN or —OH; and phenyl,optionally substituted with —SO₂NH₂; or —(X)— is O, and R⁴ is selectedfrom C₁₋₃alkyl; —CH₂SO₂C₁₋₃alkyl; and heterocyclyl selected fromtetrahydropyranyl, tetrahydrofuranyl, pyrrolidinyl, piperidinyl, andazetidinyl, wherein said heterocyclyl is optionally substituted with oneto three groups selected from —C(O)C₁₋₃alkyl, halogen, —OH, oxo andC₁₋₃alkyl; or X is (—CH₂—), and R⁴ is selected from —SO₂C₁₋₃alkyl;—C(O)N(C₁₋₃alkyl)₂; —NHC(O)R⁵ or —N(CH₃)C(O)R⁵, wherein R⁵ is selectedfrom cyclopropyl and C₁₋₃alkyl optionally substituted with one to three—F groups; —OCH₂C(O)N(C₁₋₃ alkyl)₂; —NHSO₂C₁₋₃alkyl; —S(═O)(═NH)CH₃;heterocyclyl selected from pyrrolidinyl, 1,1-dioxo[1,21-thiazine,morpholinyl and oxazolidinyl, wherein said heterocyclyl is optionallysubstituted with one to three groups selected from —C(O)C₁₋₃alkyl,halogen, —OH, oxo and C₁₋₃alkyl; and —C(O)-heterocyclyl, wherein theheterocyclyl is selected from morpholin-4-yl, pyrrolidin-1-yl andpiperidin-1-yl, optionally substituted with one or two groups selectedfrom —F and —OH; and R³ is H or C₁₋₃alkyl optionally substituted with—OH; or a salt or a stereoisomer thereof.
 3. The compound according toclaim 1, wherein: R² is —(X)—R⁴, wherein —(X)— is a bond, and R⁴ isselected from —CF₃; —CHF₂; —CH₂OH; —CH(OH)CH₃; —CH(OH)CF₃; —F; —CN;heterocyclyl selected from tetrahydropyranyl and pyrrolidinyl, whereinsaid heterocyclyl is substituted with one to three groups selected fromC₁₋₃alkyl, —F, —OH and oxo; C₃₋₆cycloalkyl, substituted with —CN or —OH;and phenyl, optionally substituted with —SO₂NH₂; and R³ is H, orC₁₋₃alkyl optionally substituted with —OH; or a salt or a stereoisomerthereof.
 4. The compound according to claim 1, wherein: R² is —(X)—R⁴,wherein —(X)— is O, and R⁴ is selected from C₁₋₃alkyl; —CH₂SO₂C₁₋₃alkyl;and heterocyclyl selected from tetrahydropyranyl, tetrahydrofuranyl,pyrrolidinyl, piperidinyl, and azetidinyl, wherein said heterocyclyl isoptionally substituted with —C(O)C₁₋₃alkyl; and R³ is H, or C₁₋₃alkyloptionally substituted with —OH; or a salt or a stereoisomer thereof. 5.The compound according to claim 1, wherein: R² is —(X)—R⁴, wherein X is(—CH₂—), and R⁴ is selected from —SO₂C₁₋₃alkyl; —C(O)N(C₁₋₃alkyl)₂;—NHC(O)R⁵ or —N(CH₃)C(O)R⁵, wherein R⁵ is selected from cyclopropyl andC₁₋₃alkyl optionally substituted with one to three —F groups;—OCH₂C(O)N(C₁₋₃ alkyl)₂; —NHSO₂C₁₋₃alkyl; —S(═O)(═NH)CH₃; heterocyclylselected from pyrrolidinyl, 1,1-dioxo[1,2]-thiazine, morpholinyl andoxazolidinyl, wherein said heterocyclyl is optionally substituted withone to two groups selected from oxo and C₁₋₃alkyl; and—C(O)-heterocyclyl, wherein the heterocyclyl is selected frommorpholin-4-yl, pyrrolidin-1-yl and piperidin-1-yl, optionallysubstituted with one or two groups selected from —F and —OH; and R³ isH, or C₁₋₃alkyl optionally substituted with —OH; or a salt or astereoisomer thereof.
 6. The compound according to claim 1, wherein: R¹is —C(O)NH₂; or a salt or a stereoisomer thereof.
 7. The compoundaccording to claim 1, wherein: R¹ is —CN; or a salt or a stereoisomerthereof.
 8. The compound according to claim 1 selected from the groupconsisting of

or a pharmaceutically acceptable salt or a stereoisomer thereof.
 9. Thecompound according to claim 8 selected from the group consisting ofcompound numbers 1, 5, 12, 29, 37, 43, 56, 61, and 62 or apharmaceutically acceptable salt or a stereoisomer thereof.
 10. Apharmaceutical composition comprising a compound according to claim 1and a pharmaceutically acceptable excipient or carrier.
 11. A method oftreating a disease or disorder that can be alleviated by inhibition ofaldosterone synthase selected from diabetic nephropathy,glomerulosclerosis, glomerulonephritis, IGA nephropathy, nephriticsyndrome, focal segmental glomerulosclerosis (FSGS), hypertension,systolic heart failure, diastolic heart failure, left ventriculardysfunction, arterial stiffness, and adrenal hyperplasia and primary andsecondary hyperaldosteronism, the method comprising administering atherapeutically effective amount of a compound according to claim 1 topatient in need thereof.
 12. The method according to claim 11, whereinthe disease or disorder is selected from diabetic nephropathy,glomerulosclerosis, glomerulonephritis, IGA nephropathy, nephriticsyndrome and focal segmental glomerulosclerosis (FSGS).
 13. The methodaccording to claim 11 wherein the disease is diabetic nephropathy.
 14. Amethod of treating a disease or disorder that can be alleviated byinhibition of aldosterone synthase selected from pulmonary arterialhypertension, Conn's syndrome, left ventricular stiffness and fibrosis,left ventricular filling abnormalities, and atherosclerosis, the methodcomprising administering a therapeutically effective amount of acompound according to claim 1 to patient in need thereof.
 15. Thecompound according to claim 1, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 16. The compoundaccording to claim 1, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 17. The compoundaccording to claim 1, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 18. The compoundaccording to claim 1, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 19. The compoundaccording to claim 1, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 20. The compoundaccording to claim 1, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 21. A pharmaceuticalcomposition comprising the compound according to claim 15, or apharmaceutically acceptable salt thereof, and at least onepharmaceutically acceptable carrier.
 22. A pharmaceutical compositioncomprising the compound according to claim 16, or a pharmaceuticallyacceptable salt thereof, and at least one pharmaceutically acceptablecarrier.
 23. A pharmaceutical composition comprising the compoundaccording to claim 17, or a pharmaceutically acceptable salt thereof,and at least one pharmaceutically acceptable carrier.
 24. Apharmaceutical composition comprising the compound according to claim18, or a pharmaceutically acceptable salt thereof, and at least onepharmaceutically acceptable carrier.
 25. A pharmaceutical compositioncomprising the compound according to claim 19, or a pharmaceuticallyacceptable salt thereof, and at least one pharmaceutically acceptablecarrier.
 26. A pharmaceutical composition comprising the compoundaccording to claim 20, or a pharmaceutically acceptable salt thereof,and at least one pharmaceutically acceptable carrier.
 27. The methodaccording to claim 11, wherein the compound of formula (I) is

or a pharmaceutically acceptable salt thereof.
 28. The method accordingto claim 11, wherein the compound of formula (I) is

or a pharmaceutically acceptable salt thereof.
 29. The method accordingto claim 11, wherein the compound of formula (I) is

or a pharmaceutically acceptable salt thereof.
 30. The method accordingto claim 11, wherein the compound of formula (I) is

or a pharmaceutically acceptable salt thereof.
 31. The method accordingto claim 11, wherein the compound of formula (I) is

or a pharmaceutically acceptable salt thereof.
 32. The method accordingto claim 11, wherein the compound of formula (I) is

or a pharmaceutically acceptable salt thereof.